KR20170002610A - Method of treating triple-negative breast cancer using thienotriazolodiazepine compound - Google Patents

Abstract

The present invention relates to a thienotriazololadia zepine compound of the formula (1) wherein R ¹ is alkyl of 1-4 carbon atoms, in combination with at least one chemotherapeutic agent selected from the group consisting of mTOR inhibitors and mitotic inhibitors and, R ² is a hydrogen atom; a halogen atom or a hydroxyl group optionally substituted with a carbon number of 1 to 4 alkyl, R ³ is a halogen atom;; a halogen atom a halogen atom, an alkyl carbon number of 1-4, the carbon number is 1 -NR ⁵ - (CH ₂ ) _m -R ⁶ wherein R ⁵ is a hydrogen atom or alkyl having from 1 to 4 carbon atoms, m is an integer from 0 to 4, , R ⁶ is a phenyl or pyrido dilim optionally substituted with a halogen atom); or _{^{-NR 7 -CO- (CH 2) n}} -R 8 ( wherein, R ⁷ is a hydrogen atom or an alkyl having 1 to 4 carbons, n is is an integer from 0-2, R ⁸ is phenyl or pyrido dilim optionally substituted with a halogen atom), R ⁴ is - (CH _{2) a} - CO-NH-R ⁹ wherein a is an integer of 1 to 4, R ⁹ is alkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, alkoxy of 1-4 carbon atoms, (CH2) _b -COOR < ₁₀ > wherein b is an integer of 1 to 4 and R < ¹⁰ > is an alkyl group having 1 to 4 carbon atoms, alkoxy having 1 to 4 carbon atoms, phenyl or pyridyl optionally substituted with an amino or hydroxyl group, Is an alkyl having from 1 to 4 carbon atoms, or a pharmaceutically acceptable salt thereof or a hydrate or solvate thereof, in a mammal comprising administering to the patient a pharmaceutically acceptable amount of tripeptide Provides a method of treating breast cancer.

Description

METHOD OF TREATING TRIPLE-NEGATIVE BREAST CANCER USING THIENOTRIAZOLODIA ZEPINE COMPOUND BACKGROUND OF THE INVENTION 1. Field of the Invention < RTI ID = 0.0 >

The present application describes a method of treating triple negative breast cancer using a thienotriazololadiazepine compound that is improved in solubility and bioavailability and can be provided in the form of a solid dispersion.

The compounds of formula (1) described herein below are characterized by the addition of acetylated histone H4 to the BRD containing family of transcription factors known as BET (bromodomains and extraterminal) proteins, including BRD2, BRD3 and BRD4 Lt; / RTI > See U.S. Published Patent Application No. 2010/0286127 A1, which is incorporated herein by reference in its entirety. The BET protein is known as a major post-regulatory factor in proliferation and differentiation, and is also described in Denis, GV "Bromodomain coactivators in cancer, obesity, type 2 diabetes, and inflammation," Discov Med 2010; 10: 489-499], a high inflammatory profile and a risk for cardiovascular disease and type 2 diabetes, as well as an increased risk of autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus It is associated with high susceptibility to Lupus. Thus, the compounds of formula (1) may be useful for the treatment of a variety of cancers, cardiovascular diseases, type 2 diabetes, and autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus.

Triple-negative breast cancer (TNBC) is an aggressive and heterogeneous subtype of clinically defined breast cancer that lacks estrogen and progesterone receptors, including human epithelial growth factor receptor 2 (HER2). A small number of therapeutic options have shown clinical benefit over cytotoxic combination therapy. Clinical trials have demonstrated that more than 50% of human breast cancers have a median O ₂ partial pressure that is much lower than normal breast tissue and are correlated with chemical resistance and radioactivity tolerance. Here, the antitumor activity of the compound (1-1) (also referred to herein as OTX015) in a normal oxygen and hypoxic environment, including a combination with an antitumor agent, was examined.

In some embodiments, the present invention provides a method of treating triple negative breast cancer using the compositions described herein.

In some embodiments, the invention is directed to a method of treating a subject in need thereof, comprising administering to a patient in need of a pharmaceutically acceptable amount of a composition comprising a solid dispersion according to any of the compositions described in Sections III, IV, V and VI described herein Lt; RTI ID = 0.0 > triple < / RTI > negative breast cancer in a mammal.

In some embodiments, the present invention provides a compound of formula (1), specifically a compound of formula (1A), for use in treating triple negative breast cancer.

In some embodiments, the present disclosure provides solid dispersions according to any of the compositions described in Sections III, IV, V, and VI described herein for use in treating triple negative breast cancer.

In some embodiments, the present invention provides a thienotriazololodiazepine compound of the formula (1), or a pharmaceutically acceptable salt or hydrate or solvate thereof, in combination with one or more chemistries selected from the group consisting of mTOR inhibitors and mitotic inhibitors Lt; RTI ID = 0.0 > triple-negative breast cancer < / RTI >

In this formula,

R ¹ is an alkyl carbon number of 1-4,

R ² is a hydrogen atom; A halogen atom; Or alkyl having from 1 to 4 carbon atoms optionally substituted with a halogen atom or a hydroxyl group,

R ³ is a halogen atom; A halogen atom, alkyl having 1 to 4 carbon atoms, alkoxy having 1 to 4 carbon atoms, or cyano; Wherein R ⁵ is a hydrogen atom or alkyl having 1 to 4 carbon atoms, m is an integer of 0 to 4, R ⁶ is phenyl or pyridyl optionally substituted with a halogen atom, -NR ⁵ - (CH ₂ ) _m -R ⁶ ); Or -NR ⁷ -CO- (CH ₂ ) _n -R ⁸ wherein R ⁷ is a hydrogen atom or alkyl having 1 to 4 carbon atoms, n is an integer of 0 to 2, R ⁸ is phenyl optionally substituted with a halogen atom Or pyridyl,

R ⁴ is - (CH ₂ ) _a -CO-NH-R ⁹ wherein a is an integer of 1 to 4, R ⁹ is alkyl having 1 to 4 carbon atoms, hydroxyalkyl having 1 to 4 carbon atoms, 1-4 alkoxy; or an alkyl carbon number of 1-4, the carbon number is 1-4 alkoxy, amino or hydroxyl group or a phenyl group optionally substituted pyrido dilim) or ^{_{- (CH 2) b -COOR 10}} ( wherein b is And R < ¹⁰ > is alkyl having 1-4 carbon atoms.

In some embodiments, Formula (1) is selected from Formula (1A), a pharmaceutically acceptable salt thereof, or a hydrate thereof:

Wherein R ¹ is C ₁ -C ₄ alkyl, R ² is C ₁ -C ₄ alkyl, a is an integer of 1-4, R ³ is C ₁ -C ₄ alkyl, C ₁ -C ₄ alkyl, -C ₄ hydroxyalkyl, C ₁ -C ₄ alkoxy, the substituents as defined in R ⁹ of the general formula (1) phenyl, or formula (1) has a substituent (s) as defined in R ^9, optionally in the ( Lt; / RTI > optionally substituted heteroaryl.

In one such embodiment, the thienotriazololidiazepine compound is formulated as a solid dispersion comprising an amorphous thienotriazololadiazepine compound and a pharmaceutically acceptable polymer.

In one embodiment, the compound of formula (1) is (i) (S) -2- [4- (4- chlorophenyl) -2,3,9- trimethyl- [1,2,4] triazolo- [4,3-a] [1,4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide or a dihydrate thereof; (ii) Synthesis of methyl (S) - {4- (3'-cyanobiphenyl-4-yl) -2,3,9-trimethyl-6H- (S) - (2,3,9-trimethyl-4- (4-phenylaminophenyl) - 1, 6H-thieno [3,2-f] [1,2,4] triazolo [4,3-a] [1,4] diazepin-6-yl} acetate; And (iv) methyl (S) - {2,3,9-trimethyl-4- [4- (3- phenylpropionylamino) phenyl] -6H- thieno [ 4] triazolo [4,3-a] [1,4] diazepin-6-yl} acetate.

In one embodiment, the thienotriazolyldiazepine compound of formula (1) is (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl-6H- thieno [3 , 2-f] [1,2,4] triazolo- [4,3- a] [1,4] diazepin-6-yl] -N- (4- hydroxyphenyl) acetamide dihydrate.

In one embodiment, the thienotriazolyldiazepine compound of formula (1) is (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl-6H- thieno [3 , 2-f] [1,2,4] triazolo- [4,3-a] [1,4] diazepin- 6- yl] -N- (4- hydroxyphenyl) acetamide.

In one embodiment, the thienotriazole rhodiazepine compound of formula (1) and a chemotherapeutic agent are administered simultaneously.

In one embodiment, the thienotriazole rhodiazepine compound of formula (1) and the chemotherapeutic agent are administered sequentially.

In one embodiment, the mTOR inhibitor is everolimus and the mitotic inhibitor is docetaxel.

In one embodiment, the thienotriazolidiazepine compound of formula (1) is formed as a solid dispersion. In one embodiment, the solid dispersion comprises an amorphous thienotriazolol diazepine compound of formula (1), or a pharmaceutically acceptable salt or hydrate thereof; And pharmaceutically acceptable polymers. In one embodiment, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). In one embodiment, the pharmaceutically acceptable polymer is hydroxypropylmethylcellulose acetate succinate (HPMCAS) and the weight ratio of thienotriazololodiazepine compound to hydroxypropylmethylcellulose acetate succinate is from 1: 3 to 1: 1 to be. In one embodiment, the solid dispersion exhibits a single glass transition temperature (Tg) inflection point in the range of about 130 캜 to about 140 캜.

In one embodiment, the solid dispersion is (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl-6H-thieno [3,2- f] ] Triazolo- [4,3-a] [1,4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide dihydrate or a pharmaceutically acceptable salt thereof or a hydrate thereof, Thienotriazololadia zepin compounds; And pharmaceutically acceptable polymers. In one embodiment, the solid dispersion is (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl-6H-thieno [3,2- f] ] Diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide dihydrate, the diffraction line associated with the crystalline thienotriazolo Lt; / RTI > shows an X-ray powder diffraction pattern that is substantially absent. In one embodiment, the pharmaceutically acceptable polymer is hydroxypropylmethylcellulose acetate succinate and the weight ratio of thienotriazololodiazepine compound to hydroxypropylmethylcellulose acetate succinate (HPMCAS) is from 1: 3 to 1: 1 . In one embodiment, the solid dispersion exhibits a single glass transition temperature (Tg) inflection point in the range of about 130 캜 to about 140 캜.

It is to be understood that any embodiment of the compounds according to formula (1) described herein may be used in any of the embodiments of the pharmaceutical compositions described herein unless otherwise stated. In addition, any compound or pharmaceutical composition described herein as an embodiment of the present invention may be used as a drug for treating triple negative breast cancer, as specifically described in the embodiments herein, unless otherwise stated.

BRIEF DESCRIPTION OF THE DRAWINGS The foregoing summary, as well as the following detailed description of the pharmaceutical compositions and methods of the present invention, including thienotriazololodiazepine formulations, will be better understood when read in conjunction with the accompanying drawings of illustrative embodiments. However, it should not be understood that the present invention is limited to the detailed schemes and means shown.
In the drawings,
Figure 1a shows the dissolution profile of a comparative formulation comprising a solid dispersion comprising 25% compound (1-1) and Eudragit L100-55;
Figure IB shows the dissolution profile of a comparative formulation comprising a solid dispersion comprising 50% Compound (1-1) and Eudragit L100-55;
Figure 1c shows the dissolution profile of an exemplary formulation comprising a solid dispersion comprising 25% Compound (1-1) and polyvinylpyrrolidone (PVP);
Figure ID shows the dissolution profile of an exemplary formulation comprising a solid dispersion comprising 50% Compound (1-1) and PVP;
Figure 1e shows the dissolution profile of an exemplary formulation comprising a solid dispersion comprising 25% compound (1-1) and PVP-vinyl acetate (PVP-VA);
Figure 1f shows the dissolution profile of an exemplary formulation comprising a solid dispersion comprising 50% Compound (1-1) and PVP-VA;
Figure 1g shows the dissolution profile of an exemplary formulation comprising a solid dispersion comprising 25% Compound (1-1) and Hypromellose Acetate Succinate (HPMCAS-M);
Figure 1h shows the dissolution profile of an exemplary formulation comprising a solid dispersion comprising 50% Compound (1-1) and HPMCAS-M;
Figure 1i shows the dissolution profile of an exemplary formulation comprising a solid dispersion comprising 25% compound (1-1) and hyphomelose phthalate (HPMCP-HP55);
Figure 1j shows the dissolution profile of an exemplary formulation comprising a solid dispersion comprising 50% Compound (1-1) and HPMCP-HP55;
Figure 2a shows the results of an in vivo screening of an exemplary formulation comprising a 25% compound (1-1) and a solid dispersion of PVP;
Figure 2b shows the results of an in vivo screening of an exemplary formulation comprising a solid dispersion of 25% Compound (1-1) and HMPCAS-M;
Figure 2c shows the results of an in vivo screening of an exemplary formulation comprising a solid dispersion of 50% Compound (1-1) and HMPCAS-M;
Figure 3 shows a powder X-ray diffraction profile of a solid dispersion of compound (1-1);
Figure 4a shows a modified differential scanning calorimetry record of a 25% compound (1-1) and a solid dispersion of PVP equilibrated under ambient conditions;
Figure 4b shows a modified differential scanning calorimetry record of the solid dispersion of 25% compound (1-1) and HMPCAS-M equilibrated under ambient conditions;
Figure 4c shows a modified differential scanning calorimetry record of a solid dispersion of 50% Compound (1-1) and HMPCAS-M equilibrated under ambient conditions;
5 is a graph of the glass transition temperature (Tg) relative to the relative humidity (RH) of the solid dispersion of 25% compound (1-1) and PVP or HMPCAS-M and 50% ;
Figure 6 shows a modified differential scanning calorimetry record of a 25% compound (1-1) and a solid dispersion of PVP equilibrated under 75% relative humidity;
Figure 7b is a graph showing the effect of the compound (1-1): 25% compound (1-1): PVP (open circular), 25% compound (1-1): HPMCAS-MG (open triangle) (Closed triangles) and 1 mg / kg intravenous dose (as inverse triangles), and a 3 mg / kg oral dose. Figure 7a shows the same data plotted in the semi-log range;
FIG. 8B shows the results of an experiment using a 25% compound (1-1): PVP (open circular), 25% compound (1-1): HPMCAS-MG (open triangle) Inverse triangles) of the compound (1-1) after oral administration of 3 mg / kg orally. Figure 8a shows the same data drawn in the semi-log range;
Figure 9 shows the powder X-ray diffraction profile of the solid dispersion of compound (1-1) in HPMCAS-MG at 0 of the stability test;
Figure 10 shows a powder X-ray diffraction profile of a solid dispersion of compound (1-1) in HPMCAS-MG after 1 month at 40 ° C and 75% relative humidity;
Figure 11 shows a powder X-ray diffraction profile of a solid dispersion of compound (1-1) in HPMCAS-MG after 2 months at 40 ° C and 75% relative humidity;
Figure 12 shows a powder X-ray diffraction profile of a solid dispersion of HPMCAS-MG compound (1-1) after 3 months at 40 < 0 > C and 75% relative humidity;
Figure 13 shows the GI50, and E _max values of HCC1937, MDA-MB-231 and MDA-MB-468 cells treated with Compound (1-1);
Figure 14a shows cell cycle steps% of G1, S, G2 / M over time in the drug-free medium and compound (1-1) of the HCC1937 cell line;
Figure 14b shows the cell cycle steps of G1, S, G2 / M over time in the drug-free medium and compound (1-1) of the MDA-MB-231 cell line;
Figure 14c shows cell cycle steps% of G1, S, G2 / M over time in the drug-free medium and compound (1-1) of the MDA-MB-468 cell line;
Figure 15a shows the basic western blot profiles of C-MYC, BRD2, BRD3, BRD4 and [beta] -tubulin in HCC1937, MDA-MB-231 and MDA-MB-468 cell lines;
Figure 15b shows the fluorescence intensities for basal levels of C-MYC in HCC1937, MDA-MB-231 and MDA-MB-468 cell lines;
Figure 15c shows the fluorescence intensities for basal levels of BRD2 in HCC1937, MDA-MB-231 and MDA-MB-468 cell lines;
Figure 15d shows the fluorescence intensities for basal levels of BRD3 in HCC1937, MDA-MB-231 and MDA-MB-468 cell lines;
15E shows the fluorescence intensity for the basal levels of BRD4 in HCC1937, MDA-MB-231 and MDA-MB-468 cell lines;
16A shows the Western blot profile of C-MYC, BRD2, BRD3, BRD4 and [beta] -tubulin in the HCC1937 cell line treated with 650 nM compound (1-1);
Figure 16b shows the Western blot profile of C-MYC, BRD2, BRD3, BRD4 and [beta] -tubulin in the MDA-MB-231 cell line treated with 75 nM compound (1-1);
Figure 16c shows the Western blot profile of C-MYC, BRD2, BRD3, BRD4 and [beta] -tubulin in the MDA-MB-468 cell line treated with 650 nM compound (1-1);
16D shows the fluorescence intensities of C-MYC when treated with 650 nM, 75 nM and 650 nM compound (1-1) for 24, 48 and 72 hours, respectively, in HCC1937, MDA-MB-231 and MDA- ;
16E shows the fluorescence intensity of BRD2 upon treatment of HCC1937, MDA-MB-231 and MDA-MB-468 cell lines with 650 nM, 75 nM, 650 nM compound (1-1) for 24, 48 and 72 hours, respectively ;
16F shows the fluorescence intensity of BRD3 upon treatment of HCC1937, MDA-MB-231 and MDA-MB-468 cell lines with 650 nM, 75 nM and 650 nM compound (1-1) for 24, 48 and 72 hours, respectively ;
Figure 16g shows the fluorescence intensity of BRD4 upon treatment of HCC1937, MDA-MB-231 and MDA-MB-468 cell lines with 650 nM, 75 nM, 650 nM compound (1-1) for 24, 48 and 72 hours, respectively ;
FIGS. 17A and 17B show the combination index values of HCC1937, MDA-MB-231 and MDA-MB-468 cell lines in which compound (1-1) was combined with beroolimus.
Figure 18a shows the tumor weights according to days after cell inoculation in the control, the compound (1-1), the berolimus and the compound (1-1) and the berolimus combination;
FIG. 18B shows the body weight according to days after cell inoculation in the control group, the compound (1-1), berolimus and the compound (1-1) and berolimus combination.
Figure 19 shows the effect of compound (1-1) on the cell cycle over time in triple negative breast cancer cell lines;
Figures 20a and 20b show the effect of compound (1-1) on BRD2 / 3/4 and c-Myc expression;
Figure 21 shows the effect of compound (1-1) in combination with everolimus (A) or docetaxel (B) at 48 hours and 72 hours in TNBC cells under normal oxygen and hypoxic conditions.

The subject matter of the present invention will now be described more fully hereinafter with reference to the accompanying drawings and embodiments, which represent representative embodiments. However, the subject matter may be embodied in various forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided to enable those skilled in the art to make and such modifications. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the relevant arts. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

I. Definition:

As used herein, the term "alkyl group" means a saturated linear or branched hydrocarbon.

The term "substituted alkyl group" means an alkyl moiety having at least one substituent which substitutes at least one carbon or hydrogen of the hydrocarbon backbone.

The term "alkenyl group ", alone or in combination with a partially unsaturated branched chain having at least one carbon-carbon double bond, whether a substituent such as" C _1-4 alkenyl (aryl) Refers to a hydrocarbon radical in which the double bond is derived by the removal of one hydrogen atom at each of two adjacent carbon atoms of the parent alkyl molecule and the radical is induced by the removal of one hydrogen atom at a single carbon atom. The atom can be oriented to a double bond in a cis (Z) or trans (E) steric form. Typical alkenyl radicals include, without limitation, ethenyl, propenyl, allyl (2-propenyl), butenyl, and the like. Examples include C _2-8 alkenyl or C _2-4 alkenyl groups.

The term "C _{(jk)" (where,} j and k is an integer indicating the carbon atoms of the given number) is alkyl, alkenyl, alkynyl, alkoxy, or means a cycloalkyl radical, or or alkyl is k carbon atoms from j ≪ / RTI > is meant an alkyl moiety of a radical represented by a covalent bond. For example, C _(1-4) means a radical containing 1, 2, 3 or 4 carbon atoms.

The term " halo "or" halogen "as used herein means F, Cl, Br, or I.

The term "pharmaceutically acceptable salts" is art-recognized and includes, for example, the relatively non-toxic, inorganic and organic acid addition salts of compounds, including inorganic or organic base addition salts Means salt.

The term "solid dispersion " as used herein means a group of solid products consisting of at least two different components, generally a hydrophilic carrier and a hydrophobic drug (active ingredient).

The term " chiral "is art-recognized and refers to molecules having non-superimposing properties of enantiomeric partners, while the term" non-chiral " means molecules that can be superimposed on their enantiomeric partners. A "prochiral molecule" is a molecule that has the potential to be converted into a chiral molecule by a specific process.

"는 단일, 이중 또는 삼중 결합일 수 있는 결합을 의미하는 데 사용된다. Symbol "

Quot; is used herein to mean a bond which may be a single, double or triple bond.

The term "enantiomer ", as used herein, and the structural formulas depicting the enantiomer are enantiomeric enantiomers, including enantiomeric excesses such as at least 10%, 25%, 50% , 75%, 90%, 95%, 98%, or 99% enantiomeric excess and an enantiomer thereof.

As used herein, the term "stereoisomer" is intended to include all geometric isomers, enantiomers or diastereomers. The present invention includes various stereoisomers of these compounds and mixtures thereof. Form isomers and rotamers of the compounds disclosed herein are also contemplated.

The term " stereoselective synthesis " as used herein means a chemical or enzymatic reaction in which a single reactant forms a heterogeneous mixture of stereoisomers during the generation of a new stereogenic center or a modification of a pre-existing one, and is well known in the art . Stereoselective synthesis involves both enantioselective and diastereomeric selective modifications. See, for example, Carreira, EM and Kvaerno, L., Classics in Stereoselective Synthesis , Wiley-VCH: Weinheim, 2009.

The term "spray drying" refers to the rapid removal of solvent from a mixture in a processor chamber where there is a strong driving force (e.g., hot dry gas or partial vacuum, or a combination thereof) for atomization of the feed suspension or solution into small droplets, . ≪ / RTI >

As used herein, the term "therapeutically effective amount" refers to an amount of a therapeutically effective amount of a thienothiazol-zoloadzepine or other pharmacologically active agent that, when administered to a patient, Or any amount of a thienotriazololodiazepine of the invention or any other pharmacologically active agent that reduces progression of the disease or disorder.

The term "about" means +/- 10%. In one embodiment, this means +/- 5%.

In the present application and in the claims below, unless otherwise required, variations such as the word " comprises ", or "comprises" or "comprising " mean inclusive of a stated integer phase or group of integers or steps But does not exclude any other integers or steps or groups of integers or steps. It should also be understood that the term "comprising"

The thienotriazololodiazepine compounds of the formula (1) described herein can be formulated as pharmaceutically acceptable polymers and solid dispersions to provide oral preparations that provide a high absorption of the pharmaceutical ingredients from the gastrointestinal tract into the circulatory system . In one embodiment, the pharmaceutically acceptable polymer is hypromellose acetate succinate (also referred to as hydroxypropylmethylcellulose acetate succinate or HPMCAS). In one embodiment, the pharmaceutically acceptable polymer is polyvinylpyrrolidone (PVP).

In some embodiments, hydroxypropylmethylcellulose acetate succinate (HPMCAS) is prepared by mixing 9% acetyl / 11% succinoyl (e.g., having an average particle size of 5 쨉 m (i.e., HPMCAS-MF, (E.g., HPMCAS-HF, saphale grade) with 12% acetyl / 6% succinonyl (e.g., HPMCAS-HF, HPMCAS with 1 mm Or HPMCAS having an average particle size of 1 mm (i.e., HPMCAS-HG, granule grade), and 8% acetyl / 15% -LF, seed grade) or HPMCAS with an average particle size of 1 mm (i.e., HPMCAS-LG, granular grade).

In some embodiments, the polyvinylpyrrolidone has a weight average molecular weight of about 2,500 (Kollidon 12 PF, weight average molecular weight of 2,000 to 3,000), about 9,000 (Kollidon 17 PF, weight average molecular weight of 7,000 to 11,000), about 25,000 Molecular weight of about 50,000 (Kollidon (R) 30, weight average molecular weight of 44,000 to 54,000), or about 1,250,000 (Kollidon 90 or Kollidon 90F, 1,000,000 to 1,500,000 weight average molecular weight) Lt; / RTI >

II. Treatment method

In some embodiments, the present invention provides a method of treating triple negative breast cancer using the compositions described herein.

In some embodiments, the invention provides a method comprising administering to a patient in need of a pharmaceutically acceptable amount of a composition comprising a solid dispersion according to any of the compositions described in Sections III, IV, V and VI described herein Lt; RTI ID = 0.0 > triple-negative breast < / RTI >

In some embodiments, the method includes administering to a patient in need of a pharmaceutically acceptable amount of a composition comprising a pharmaceutical preparation according to any of the compositions described in Sections III, IV, V and VI described herein Lt; RTI ID = 0.0 > triple-negative < / RTI > breast cancer in a mammal.

In some embodiments, the present invention provides a compound of formula (1), specifically a compound of formula (1A), for use in treating triple negative breast cancer.

In some embodiments, the present disclosure provides solid dispersions according to any of the compositions described in Sections III, IV, V, and VI described herein for use in treating triple negative breast cancer.

In some embodiments, the method of treating triple negative breast cancer comprises administering to a subject in need thereof a therapeutically effective amount of a Thienotriazole derivative of formula (1), including any salt, isomer, enantiomer, racemate, hydrate, solvate, metabolite, Zolodia zepin compounds are used:

In this formula,

R ¹ is an alkyl carbon number of 1-4,

R ² is a hydrogen atom; A halogen atom; Or alkyl having from 1 to 4 carbon atoms optionally substituted with a halogen atom or a hydroxyl group,

R ³ is a halogen atom; A halogen atom, alkyl having 1 to 4 carbon atoms, alkoxy having 1 to 4 carbon atoms, or cyano; _{^{-NR 5 - (CH 2) m}} -R 6 ( wherein, R ⁵ is a hydrogen atom or an alkyl having 1 to 4 carbons, m is an integer from 0-4, R ⁶ is a halogen atom, an optionally substituted phenyl or flutes Dylem); Or -NR ⁷ -CO- (CH ₂ ) _n -R ⁸ wherein R ⁷ is a hydrogen atom or alkyl having 1 to 4 carbon atoms, n is an integer from 0 to 2, and R ⁸ is optionally substituted with a halogen atom Phenyl or pyridyl,

R ⁴ is - (CH ₂ ) _a -CO-NH-R ⁹ (wherein a is an integer of 1 to 4, R ⁹ is alkyl having 1 to 4 carbon atoms, hydroxyalkyl having 1 to 4 carbon atoms, the 1-4 alkoxy; or a 1-4 carbon atoms alkyl, 1-4 carbon atoms alkoxy, amino or hydroxyl group or a phenyl group optionally substituted pyrido dilim) or ^{_{- (CH 2) b -COOR 10}} ( wherein b is an integer of 1-4, and R < ¹⁰ > is alkyl having 1-4 carbon atoms.

In some embodiments, Formula (1) is selected from a compound of Formula (1A), a pharmaceutically acceptable salt thereof or a hydrate thereof:

Wherein R ¹ is C ₁ -C ₄ alkyl, R ² is C ₁ -C ₄ alkyl, a is an integer of 1-4, R ³ is C ₁ -C ₄ alkyl, C ₁ -C ₄ alkyl, -C ₄ hydroxyalkyl, C ₁ -C ₄ alkoxy, the substituents as defined in R ⁹ of the general formula (1) phenyl, or formula (1) has a substituent (s) as defined in R ^9, optionally in the ( Lt; / RTI > optionally substituted heteroaryl.

In one such embodiment, the thienotriazololidiazepine compound is formulated as a solid dispersion comprising an amorphous thienotriazololadiazepine compound and a pharmaceutically acceptable polymer.

In the present invention, "treating" or "treating" is intended to include, for example, triple negative breast cancer or to alleviate symptoms, to prevent the onset of triple negative breast cancer or symptoms, or to restore the onset condition of triple negative breast cancer Refers to the administration of, or such administration of the active ingredient of the invention to an individual (patient) diagnosed by a physician as having a risk of developing a triple negative breast cancer or a triple negative breast cancer.

III. Thienotriazolol idia zepin compound:

In one embodiment, the thienotriazolol diazepine compounds used in the formulations of the present invention include any salts, isomers, enantiomers, racemates, hydrates, solvates, metabolites, and polymorphs thereof, (1): < EMI ID =

In this formula,

R ¹ is an alkyl carbon number of 1-4,

R ² is a hydrogen atom; A halogen atom; Or alkyl having from 1 to 4 carbon atoms optionally substituted with a halogen atom or a hydroxyl group,

R ³ is a halogen atom; A halogen atom, alkyl having 1 to 4 carbon atoms, alkoxy having 1 to 4 carbon atoms, or cyano; Wherein R ⁵ is a hydrogen atom or alkyl having 1 to 4 carbon atoms, m is an integer of 0 to 4, R ⁶ is phenyl or pyridyl optionally substituted with a halogen atom, -NR ⁵ - (CH ₂ ) _m -R ⁶ ); Or -NR ⁷ -CO- (CH ₂ ) _n -R ⁸ wherein R ⁷ is a hydrogen atom or alkyl having 1 to 4 carbon atoms, n is an integer of 0 to 2, R ⁸ is phenyl optionally substituted with a halogen atom Or pyridyl,

R ⁴ is - (CH ₂ ) _a -CO-NH-R ⁹ wherein a is an integer of 1 to 4, R ⁹ is alkyl having 1 to 4 carbon atoms, hydroxyalkyl having 1 to 4 carbon atoms, 1-4 alkoxy; or an alkyl carbon number of 1-4, the carbon number is 1-4 alkoxy, amino or hydroxyl group or a phenyl group optionally substituted pyrido dilim) or ^{_{- (CH 2) b -COOR 10}} ( wherein b is And R < ¹⁰ > is alkyl having 1-4 carbon atoms.

In one embodiment, suitable alkyl groups include linear or branched akyl radicals containing from one carbon atom to four carbon atoms. In one embodiment, suitable alkyl groups include linear or branched akyl radicals containing from 1 to 3 carbon atoms. In one embodiment, suitable alkyl groups include linear or branched akyl radicals containing from 1 to 2 carbon atoms. In one embodiment, exemplary alkyl radicals include, without limitation, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl. In one embodiment, exemplary alkyl groups include, without limitation, methyl, ethyl, propyl, isopropyl, 2-methyl-1-propyl, and 2-methyl-2-propyl.

In some embodiments, the present invention provides pharmaceutically acceptable salts, hydrates, solvates, and isotopically labeled forms of the thienotriazololodiazepine compounds described herein. In one embodiment, the pharmaceutically acceptable salts of the thienotriazoloidazepine compounds include inorganic acids and formed acid addition salts. In one embodiment, the pharmaceutically acceptable inorganic acid addition salts of thienotriazololidiazepines include salts of hydrochloric acid, bromic acid, iodic acid, phosphoric acid, metaphosphoric acid, nitric acid, and sulfuric acid. In one embodiment, the pharmaceutically acceptable salts of the thienothiazololidiazepine compounds include organic acids and formed acid addition salts. In one embodiment, the pharmaceutically acceptable organic acid addition salts of thienothiazololidiazepines are selected from the group consisting of tartaric acid, acetic acid, trifluoroacetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, formic acid, propionic acid, glycolic acid, gluconic acid , Maleic acid, succinic acid, camphorsulfonic acid, isothionic acid, mucic acid, gentisic acid, isonicotinic acid, saccharic acid, glucuronic acid, furoic acid, glutamic acid, ascorbic acid, anthranilic acid, salicylic acid, phenylacetic acid, mandelic acid , Methanesulfonic acid, ethanesulfonic acid, pantothenic acid, stearic acid, sulfinic acid, alginic acid, galacturonic acid, and arylsulfonic acids such as benzenesulfonic acid and salts of 4-methylbenzenesulfonic acid .

The present invention provides the pharmaceutically acceptable isotopically labeled forms of the thienotriazoloadiazepine compounds described herein wherein one or more atoms have the same atomic number but the atomic mass or mass number generally refers to atoms found in nature Mass or mass number. Examples of isotopes suitable for inclusion in the thienotriazoloidazepine compounds include isotopes of hydrogen, such as ² H and ³ H, carbon isotopes such as ¹¹ C, ¹³ C and ¹⁴ C, chlorine For example, ³⁶ Cl, an isotope of fluorine such as ¹⁸ F, an isotope of iodine such as ¹²³ I and ¹²⁵ I, an isotope of nitrogen such as ¹³ N and < RTI ID = 0.0 > ¹⁵ N, isotopes of oxygen such as ¹⁵ O, ¹⁷ O and ¹⁸ O, and isotopes of sulfur, for example, ³⁵ S. The isotopically labeled forms of the thienotriazololadia zepine compounds can be prepared by conventional techniques generally known to those skilled in the art.

Some isotopically labeled forms of the compounds of formula (1), for example those into which radioactive isotopes are incorporated, are useful in drug and / or substrate tissue distribution studies. Radioactive isotopes tritium ( ³ H) and carbon-14 ( ¹⁴ C) are particularly useful for that purpose in that they are easy to introduce and ready to use. Substitution with heavier isotopes such as deuterium ( ² H) may provide for certain metabolic stability, such as increased in vivo half-life or decreased dosage requirements, which may be desirable in some circumstances. Substitution with positron emitting isotopes such as ¹¹ C, ¹⁸ F, ¹⁵ O, and ¹³ N can be used in positron emission tomography (PET) studies to investigate substrate receptor occupancy.

In some embodiments, the thienotriazolyldiazepine compounds disclosed herein may exist in unsolvated forms, including solvated forms with pharmaceutically acceptable solvents. Those skilled in the art will appreciate that the solvate is a complex of various stoichiometries formed by the solute (in this case, the thienotriazolol diazepine compound described herein) and the solvent. It is preferred that such a solvent does not interfere with the biological activity of the solute (thienotriazolol diazepine compound). Examples of suitable solvents for solvate formation include, without limitation, water, methanol, dimethylsulfoxide, ethanol and acetic acid. Suitably used solvents are pharmaceutically acceptable solvents. A suitable solvent is water. In one embodiment, the pharmaceutically acceptable solvates of the thienothiazolazolidinedine compounds described herein are selected from the group consisting of ethanol solvates, isopropanol solvates, dioxolane solvates, tetrahydrofuran solvates, dimethylsulfoxide solvates, butanol solvate, 2-butanol solvate, dioxolane solvate, 1,3-dimethyl-3,4,5,6-tetrahydro-2 (1H) -pyrimidinone ("DMPU") solvate, A solvate of 1,3-dimethylimidazolidinone ("DMI"), and a solvate of 1,3-dimethylimidazolidinone ("DMP"), or mixtures thereof.

In some embodiments, the thienotriazololidiazepine compounds described herein contain one or more chiral centers and / or double bonds and can therefore exist as geometric isomers, enantiomers or diastereomers. The enantiomers and diastereoisomers of thienotriazoloadia zephin compounds are referred to as "R " moieties to uniquely identify each stereocenter (also sometimes referred to as the chiral center) by including the descriptor in its systematic name, Can be displayed in accordance with the Cahn-Ingold-Prelog convention, designated as the "S " descriptor, and each carbon-carbon double bond designated as E or Z (to indicate the geometric isomers) .

In some embodiments, the thienotriazolyldiazepine compounds described herein can exist as racemic mixtures, or racemates, comprising an equimolar amount of the left and right enantiomers of the chiral molecule. These racemic mixtures may be denoted by prefixes (±) - or dl-, representing an equal (1: 1) mixture of primary and left-sided isomers. In addition, the prefixes rac - (or racem -) or the symbols RS and SR can be used to denote racemic mixtures.

는 단일, 이중 또는 삼중 결합일 수 있는 결합을 표시하는 데 사용될 수 있다. 탄소-탄소 이중 결합 주변 치환기는 "Z" 또는 "E" 입체형태로서 표시되는데 용어 "Z" 및 "E"는 IUPAC 표준에 따라 사용된다. 달리 특정하지 않으면, 이중 결합을 묘사하는 구조는 "E" 및 "Z" 이성질체 둘 모두를 포함한다. 탄소-탄소 이중 결합 주변 치환기는 대안적으로 "cis" 또는 "trans"로 표시되는데, "cis"는 이중 결합의 동일 면 상의 치환기를 의미하고, "trans"는 이중 결합의 반대면 상의 치환기를 의미한다. 탄소환 고리 주변 치환기의 배열을 또한 "cis" 또는 "trans"로서 표시할 수 있다. 용어 "cis"는 고리 평면의 동일 면 상의 치환기를 의미하고 용어 "trans"는 고리 평면의 반대면 상의 치환기를 의미한다. 치환기가 고리 평면의 동일면 및 반대면 둘 모두에 배치된 화합물의 혼합물을 "cis/trans" 또는 "Z/E"라고 표시한다. Geometric isomers, either by arrangement of carbon-carbon double bond peripheral substituents or by arrangement of cycloalkyl or heterocyclic ring vicinal substituents, may also be present in the compounds of the present invention. In some embodiments,

May be used to indicate a bond that may be a single, double or triple bond. The carbon-carbon double bond surrounding substituents are represented as " Z " or " E " stereochemical forms, the terms " Z " and " E " Unless otherwise specified, structures describing the double bonds include both "E" and "Z" isomers. Carbon-carbon double bond peripheral substituents Alternatively, there is shown as "cis" or "trans", "cis" it is "trans" represents substituents on the same side of the double bond, and means a substituent on the reverse of the double bond plane do. The arrangement of substituents around the carbocyclic ring can also be denoted as " cis " or " trans . &Quot; The term " cis " means a substituent on the same side of the ring plane and the term " trans " means a substituent on the opposite side of the ring plane. Quot ; cis / trans "or" Z / E ", where the substituent is arranged on both the same and opposite sides of the ring plane.

In some embodiments, the thienotriazole zoloidzepine compounds disclosed herein may exist in single or multiple crystalline forms or polymorphs. In one embodiment, the thienothiazolazolidiazepine compounds disclosed herein comprise an amorphous form thereof. In one embodiment, the thienotriazolyldiazepine compounds disclosed herein comprise a single polymorph thereof. In another embodiment, the thienothiazolazolidiazepine compounds disclosed herein comprise a mixture of polymorphs thereof. In another embodiment, the compound is in crystalline form.

In some embodiments, the thienotriazolyldiazepine compounds disclosed herein may exist as a single enantiomer or as an enantiomerically enriched form. In one embodiment, the thienotriazolyldiazepine compounds disclosed herein are present in an enantiomeric excess of greater than 80%. In one embodiment, the thienotriazololodia zepine compounds disclosed herein are present in an enantiomeric excess of greater than 90%. In one embodiment, the thienotriazolyldiazepine compounds disclosed herein are present in an enantiomeric excess of greater than 98%. In one embodiment, the thienotriazololodia zepine compounds disclosed herein are present in an enantiomeric excess of greater than 99%. In some embodiments, the thienothiazololidiazepine compounds disclosed herein are at least 10%, at least 25%, at least 50%, at least 75%, at least 90%, at least 95%, at least 98% Isomer excess ratio. ≪ / RTI >

In the enantiomeric pair, the enantiomeric excess (ee) of the enantiomer E1 to the enantiomer E2 can be calculated using the following equation (1):

식 (1)

Equation (1)

The relative amounts of E1 and E2 can be determined by chiral high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) or any other suitable method. In some embodiments, the purity of the enantiomeric compound refers to the amount of enantiomers E1 and E2 relative to the amount of other material, which may clearly include byproducts and / or unreacted reactants or reagents.

In some embodiments, the thienotriazole rhodiazepine compounds of formula (1) include, without limitation, thienothiazolazolidiazepine compounds (1-1) to (1-18) listed in Table A below.

Table A : Exemplary compounds that can be used in the formulations described herein:

In some embodiments, the thienotriazolyldiazepine compound of formula (1) is (i) (S) -2- [4- (4-chlorophenyl) -2,3,9- trimethyl-6H-thieno [ 3,2-f] [1,2,4] triazolo- [4,3- a] [1,4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide or its dihydrate (Ii) methyl (S) - {4- (3'-cyanobiphenyl-4-yl) -2,3,9-trimethyl-6H-thieno [3,2- 4] triazolo [4,3-a] [1,4] diazepin-6-yl} acetate, (iii) methyl (S) - {2,3,9- ) -6H-thieno [3,2-f] [1,2,4] triazolo [4,3-a] [1,4] diazepin-6-yl} acetate; And (iv) methyl (S) - {2,3,9-trimethyl-4- [4- (3- phenylpropionylamino) phenyl] -6H- thieno [ 4] triazolo [4,3-a] [1,4] diazepin-6-yl} acetate.

In some embodiments, the thienotriazolyldiazepine compound of formula (1) is (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl- -f] [1,2,4] triazolo [4,3-a] [1,4] diazepin-6-yl] -N- (4- hydroxyphenyl) acetamide.

Exemplary mTOR (mammalian target of rapamycin) inhibitors for use in combination with the thienotriazoloidazepines of formula (1) in the methods of the present invention include, without limitation, the mTOR inhibitors listed in Table B below.

Table B : Exemplary mTOR inhibitor compounds that can be used in combination with the thienothiazololidiazepines of formula (1)

Exemplary mitotic inhibitors for use in combination with the thienothiazololidiazepines of formula (1) in the methods of the present invention include, without limitation, the mitotic inhibitors listed in Table C below.

Table C: Exemplary mitotic inhibitor compounds that can be used in combination with the thienothiazololidiazepines of formula (1):

IV. Formulation:

The compounds of formula (1) are particularly useful for the treatment of a wide variety of conditions, including, but not limited to, specific problems of drug bioavailability and variability of the dose response between patients and patients, and indispensable development of unconventional formulations, Suggests highly specific differences in manufacturing.

Previously, the compounds of formula (1) have been used to treat oral inflammatory bowel disease, such as ulcerative colitis and Crohn's disease, to provide an oral formulation which preferentially releases the pharmaceutical ingredient in the lower colon. The carrier ethyl acrylate-methyl methacrylate- It has been found that it can be formulated as a solid dispersion with trimethylammonioethyl methacrylate chloride copolymer (Eudragit RS, Rohm) (US patent application publication no. 20090012064 A1, published January 8, 2009) ). In inflammatory bowel disease, drug efflux in lesions and its direct action on inflammatory lesions have been confirmed through a variety of experiments, including animal experiments, which are more important than drug absorption from the gastrointestinal tract into the circulatory system.

Solvates, racemates, enantiomers, isomers and isotopically labeled forms, including thienotriazololodiazepine compounds, pharmaceutically acceptable salts, hydrates thereof, according to formula (1) The present invention has unexpectedly discovered that it can be formulated as a pharmaceutically acceptable polymer and a solid dispersion to provide an oral preparation that provides a high absorption of the pharmaceutical ingredient from the gastrointestinal tract into the circulatory system for treating the disease. Experiments in dogs and humans have confirmed the high oral bioavailability of these solid dispersions compared to previously developed Eudragit solid dispersion formulations for the treatment of inflammatory bowel disease.

Solid dispersions are strategies for improving oral bioavailability of non-water soluble drugs.

As used herein, the term "solid dispersion" means a solid product group comprising at least two different components, generally a hydrophilic carrier and a hydrophobic drug, a thienotriazololidiazepine compound according to formula (1). Based on the molecular arrangement of the drug in the dispersion, six different types of solid dispersions can be distinguished. Typically, solid dispersions are classified as simple eutectic mixtures, solid solutions, glass solutions and suspensions, and amorphous precipitates in crystalline carriers. Also, certain combinations can be encountered in the same sample, for example, some molecules are present as aggregates while others are molecularly dispersed.

In one embodiment, the thienotriazoloidazepine compound according to formula (1) can be molecularly dispersed as amorphous particles (clusters). In another embodiment, the thienotriazololidiazepine compound according to formula (1) may be dispersed as crystalline particles. In one embodiment, the carrier may be crystalline. In other embodiments, the carrier may be amorphous.

In one embodiment, the present invention provides a pharmaceutical composition comprising a thienotriazolol diazepine compound according to formula (1), or a pharmaceutically acceptable salt, hydrate, solvate, racemate, enantiomer, isomer, There is provided a pharmaceutical composition comprising a solid dispersion in labeled form. In one embodiment, the pharmaceutically acceptable polymer is hypromellose acetate succinate (also referred to as hydroxypropylmethylcellulose acetate succinate or HPMCAS). In one embodiment, the dispersion is a thienotriazolol diazepine compound to hydroxypropylmethyl cellulose acetate succinate (HPMCAS) weight ratio of 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazololadiazepine compound is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 130 캜 to 140 캜. In other such embodiments, a single Tg occurs at about 135 占 폚. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). For purposes of this application, "substantially free" means that there is no diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline Tienothiazolodiazepine of formula (1).

In one embodiment, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable salt, solvate, racemate, enantiomer, pharmaceutically acceptable salt, solvate, hydrate or solvate thereof, including a thienotriazololodiazepine compound of formula (I) Isomer, or isotope labeled form of the solid dispersion. In one embodiment, the pharmaceutically acceptable polymer is polyvinylpyrrolidone (also known as povidone or PVP). In one embodiment, the dispersion has a thienothiazolazolidiazepine compound to PVP weight ratio of 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazololadiazepine compound is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 175 캜 to about 185 캜. In other such embodiments, a single Tg occurs at about 179 < 0 > C. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). For purposes of this application, "substantially free" means that there is no diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline Tienothiazolodiazepine of formula (1).

In one embodiment, the pharmaceutical composition of the present invention comprises a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including an amorphous form of the thienotriazololadiazepine compound of formula (I) or a pharmaceutically acceptable salt, , Or a solid dispersion of isotopically labeled forms and pharmaceutically acceptable polymers. In one embodiment, the pharmaceutically acceptable polymer is hypromellose acetate succinate. In one embodiment, the weight ratio of thienotriazololadia zephin compound of formula (1) to heptamelose acetate succinate ranges from 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazololadiazepine compound is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 130 캜 to 140 캜. In other such embodiments, a single Tg occurs at about 135 占 폚. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). For purposes of this application, "substantially free" means that there is no diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazololidiazepine compound of formula (1).

In one embodiment, the pharmaceutical composition of the present invention comprises a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including an amorphous form of the thienotriazololadiazepine compound of formula (I) or a pharmaceutically acceptable salt, , Or a solid dispersion of isotopically labeled forms and pharmaceutically acceptable polymers. In one embodiment, the pharmaceutically acceptable polymer is polyvinylpyrrolidone. In one embodiment, the weight ratio of thienothiazolazolidinazine compound of formula (1) to polyvinylpyrrolidone ranges from 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazololadiazepine compound is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 175 캜 to about 185 캜. In other such embodiments, a single Tg occurs at about 179 < 0 > C. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). For the purposes of this application, "substantially free" means the absence of a diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazololidiazepine compound of formula (1).

In one embodiment, the pharmaceutical compositions of the present invention comprise a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including crystalline forms of the thienotriazololadiazepine compound of formula (1) , Or a solid dispersion of isotopically labeled forms and pharmaceutically acceptable polymers. In one embodiment, the pharmaceutically acceptable polymer is hypromellose acetate succinate. In one embodiment, the weight ratio of thienotriazololadia zephin compound of formula (1) to heptamelose acetate succinate ranges from 1: 3 to 1: 1.

In one embodiment, the pharmaceutical compositions of the present invention comprise a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including crystalline forms of the thienotriazololadiazepine compound of formula (1) , Or a solid dispersion of isotopically labeled forms and pharmaceutically acceptable polymers. In one embodiment, the pharmaceutically acceptable polymer is polyvinylpyrrolidone. In one embodiment, the weight ratio of thienothiazolazolidinazine compound of formula (1) to polyvinylpyrrolidone ranges from 1: 3 to 1: 1.

In some embodiments, a pharmaceutical composition comprising a solid dispersion is prepared by spray drying.

In one embodiment, the pharmaceutical compositions of the present invention comprise a solvate, racemate, enantiomer, isomer or isotope of a thienotriazolol diazepine compound of formula (I), or a pharmaceutically acceptable salt, hydrate thereof, A spray-dried solid dispersion of the labeled form and a pharmaceutically acceptable polymer. In one embodiment, the pharmaceutically acceptable polymer is hypromellose acetate succinate. In one embodiment, the weight ratio of compound (1) to heptamelose acetate succinate ranges from 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazololadiazepine compound is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 130 캜 to 140 캜. In other such embodiments, a single Tg occurs at about 135 占 폚. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). For the purposes of this application, "substantially free" means the absence of a diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazololidiazepine compound of formula (1).

In one embodiment, the pharmaceutical compositions of the present invention comprise a solvate, racemate, enantiomer, isomer or isotope of a thienotriazolol diazepine compound of formula (I), or a pharmaceutically acceptable salt, hydrate thereof, A spray-dried solid dispersion of the labeled form and a pharmaceutically acceptable polymer. In one embodiment, the pharmaceutically acceptable polymer is polyvinylpyrrolidone. In one embodiment, the weight ratio of compound (1) to polyvinylpyrrolidone ranges from 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazololadiazepine compound is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 175 캜 to 185 캜. In other such embodiments, a single Tg occurs at about 179 < 0 > C. In some of these embodiments, the solid dispersion is exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). For purposes of this application, "substantially free" means the absence of a diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazoloadienepine compound of formula (1).

In one embodiment, the pharmaceutical composition of the present invention comprises a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including an amorphous form of the thienotriazololadiazepine compound of formula (I) or a pharmaceutically acceptable salt, Or isotopically labeled forms and spray-dried solid dispersions of pharmaceutically acceptable polymers. In one embodiment, the pharmaceutically acceptable polymer is hypromellose acetate succinate. In one embodiment, the weight ratio of thienotriazololadia zephin compound of formula (1) to heptamelose acetate succinate ranges from 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazololadiazepine compound is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 130 캜 to 140 캜. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In other such embodiments, a single Tg occurs at about 135 占 폚. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). For the purposes of this application, "substantially free" means the absence of a diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazoloidazepine compound of formula (1).

In one embodiment, the pharmaceutical composition of the present invention comprises a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including an amorphous form of the thienotriazololadiazepine compound of formula (I) or a pharmaceutically acceptable salt, Or isotopically labeled forms and spray-dried solid dispersions of pharmaceutically acceptable polymers. In one embodiment, the pharmaceutically acceptable polymer is polyvinylpyrrolidone. In one embodiment, the weight ratio of thienothiazolazolidinazine compound of formula (1) to polyvinylpyrrolidone ranges from 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazolol diazepine is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 175 캜 to 185 캜. In other such embodiments, a single Tg occurs at about 179 < 0 > C. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazolodiazepine compound of formula (1). For the purposes of this application, "substantially free" means the absence of a diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazololidiazepine compound of formula (1).

In one embodiment, the pharmaceutical compositions of the present invention comprise a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including crystalline forms of the thienotriazololadiazepine compound of formula (1) Or isotopically labeled forms and spray-dried solid dispersions of pharmaceutically acceptable polymers. In one embodiment, the pharmaceutically acceptable polymer is hypromellose acetate succinate. In one embodiment, the weight ratio of thienotriazololadia zephin compound of formula (1) to heptamelose acetate succinate ranges from 1: 3 to 1: 1.

In one embodiment, the pharmaceutical compositions of the present invention comprise a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including crystalline forms of the thienotriazololadiazepine compound of formula (1) Or isotopically labeled forms and spray-dried solid dispersions of pharmaceutically acceptable polymers. In one embodiment, the pharmaceutically acceptable polymer is polyvinylpyrrolidone. In one embodiment, the weight ratio of thienothiazolazolidinazine compound of formula (1) to polyvinylpyrrolidone ranges from 1: 3 to 1: 1.

In another preferred embodiment, the present invention provides a process for the preparation of 2- [(6S) -4- (4-chlorophenyl) -2,3,9-trimethyl-6H- thienol [ 4] triazolo [4,3-a] [1,4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide dihydrate, the compound (1-1) Solvates, racemates, enantiomers, isomers, or isotopically labeled forms, including pharmaceutically acceptable salts, hydrates, and solid dispersions of pharmaceutically acceptable polymers.

In one embodiment, the pharmaceutically acceptable polymer is HPMCAS. In one embodiment, the dispersion has compound (1-1) and HPMCAS in a weight ratio of 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazolol diazepine is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In one embodiment, the solid dispersion is spray dried. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 130 캜 to 140 캜. In other such embodiments, a single Tg occurs at about 135 占 폚. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazololidiazepine compound (1-1). For the purposes of this application, "substantially free" means the absence of a diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazololadiazepine compound (1-1).

In another embodiment, the pharmaceutical composition comprises a solvate, racemate, enantiomer, isomer, or isotope labeled form, including compound (I-1) or a pharmaceutically acceptable salt, hydrate thereof; And a solid dispersion of a pharmaceutically acceptable polymer. In one embodiment, the pharmaceutically acceptable polymer is PVP. In one embodiment, the dispersion has compound (1-1) and PVP in a weight ratio of 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazolol diazepine is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In one embodiment, the solid dispersion is spray dried. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 175 캜 to 185 캜. In other such embodiments, a single Tg occurs at about 179 < 0 > C. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with crystalline thienothiazolodiazepine compounds. For the purposes of this application, "substantially free" means the absence of a diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazololadiazepine compound (1-1).

In one embodiment, the pharmaceutical composition of the present invention comprises a pharmaceutically acceptable salt, hydrate, solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, in the amorphous form of the thienotriazololodiazepine compound (1-1) Or isotopically labeled form; And a solid dispersion of a pharmaceutically acceptable polymer. In one embodiment, the pharmaceutically acceptable polymer is HPMCAS. In one embodiment, the dispersion has Compound (1-1) and HPMCAS in a weight ratio of 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazolol diazepine is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In one embodiment, the solid dispersion is spray dried. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 130 캜 to 140 캜. In other such embodiments, a single Tg occurs at about 135 占 폚. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazololidiazepine compound (1-1). For this purpose, "substantially free" means the absence of a diffraction line on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienothiazololidiazepine compound (1-1).

In one embodiment, the pharmaceutical composition of the present invention comprises a pharmaceutically acceptable salt, hydrate, solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, in the amorphous form of the thienotriazololodiazepine compound (1-1) Or isotopically labeled form; And a solid dispersion of a pharmaceutically acceptable polymer. In one embodiment, the pharmaceutically acceptable polymer is PVP. In one embodiment, the dispersion has compound (1-1) and PVP in a weight ratio of 1: 3 to 1: 1. In one embodiment, at least a portion of the thienotriazolol diazepine is homogeneously dispersed throughout the solid dispersion. In another embodiment, the thienotriazolidiazepine compound is homogeneously dispersed throughout the solid dispersion. In one embodiment, the solid dispersion is spray dried. In some embodiments, the solid dispersion exhibits a single inflection over the glass transition temperature (Tg). In some embodiments, a single Tg occurs at 175 캜 to 185 캜. In other such embodiments, a single Tg occurs at about 189 ° C. In some of these embodiments, the solid dispersion was exposed to 75% relative humidity at 40 DEG C for at least one month. In some embodiments, the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline thienothiazololidiazepine compound (1-1). For the purposes of this application, the absence of the diffraction line means on the amorphous halo, at about 21 ° 2-theta, associated with the crystalline thienotriazololadiazepine compound (1-1).

In one embodiment, the pharmaceutical compositions of the present invention comprise a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including a crystalline form of a thienotriazololadiazepine compound (1-1) or a pharmaceutically acceptable salt, Or isotopically labeled form; And a solid dispersion of a pharmaceutically acceptable polymer. In one embodiment, the pharmaceutically acceptable polymer is HPMCAS. In one embodiment, the dispersion has Compound (1-1) and HPMCAS in a weight ratio of 1: 3 to 1: 1. In one embodiment, the solid dispersion is spray dried.

In one embodiment, the pharmaceutical compositions of the present invention comprise a solvate, racemate, enantiomer, isomer, or pharmaceutically acceptable salt thereof, including a crystalline form of a thienotriazololadiazepine compound (1-1) or a pharmaceutically acceptable salt, Or isotopically labeled form; And a solid dispersion of a pharmaceutically acceptable polymer. In one embodiment, the pharmaceutically acceptable polymer is PVP. In one embodiment, the dispersion has compound (1-1) and PVP in a weight ratio of 1: 3 to 1: 1. In one embodiment, the solid dispersion is spray dried.

The solid dispersions of the present invention described herein exhibit particularly advantageous properties when administered orally. Examples of advantageous properties of solid dispersions include, without limitation, consistent and high levels of bioavailability when administered by standard bioavailability attempts in animals or humans. The solid dispersion of the present invention comprises a solid dispersion comprising a thienotriazolol diazepine compound of formula (1) and a polymer and additives. In some embodiments, the solid dispersion is prepared by dissolving the thienotriazololidiazepine compound of formula (1) and the additive of the formula (1) in water and in most aqueous media to a negligible degree The absorption of the thienotriazololadiazepine compound of the formula (1) into the bloodstream which can not be obtained simply by mixing can be achieved. The bioavailability of the thienotriazololodiazepine compound of formula (1) or thienotriazololodiazepine compound (1-1) can be determined using various in vitro and / or in vivo experiments. In vivo experiments can be performed using, for example, rats, dogs or humans.

The bioavailability can be calculated from the thienotriazololadia zepin compound or the thienotriazololadiazepine compound (1-1) of the formula (1) according to the ordinate (Y-axis) with respect to the time along the abscissa (X- (AUC) obtained by grazing the serum or plasma concentration of the test compound. Subsequently, the AUC value of the thienotriazoloadiazepine compound of formula (1) or the thienotriazololodiazepine compound (1-1) from the solid dispersion is determined by measuring the AUC value of the crystalline (1) Is compared with the AUC value of the notriazololodiazepine compound or the crystalline thienotriazololodiazepine compound (1-1). In some embodiments, the solid dispersion is selected from a curve selected from at least 0.4 times, 0.5 times, 0.6 times, 0.8 times, 1.0 times the corresponding AUC value provided by the control composition administered intravenously to the dog, (AUC), wherein the control composition comprises an equivalent amount of a crystalline thienothiazololidiazepine compound of Formula (1).

Bioavailability can be measured in vitro by simulating the pH value of gastric and intestinal environment. This measurement is carried out by suspending a solid dispersion of the thienotriazololodiazepine compound of the formula (1) or the thienotriazololodiazepine compound (1-1) in an aqueous in vitro test medium having a pH of 1.0 to 2.0 And the pH is then adjusted to pH 5.0-7.0 in the control in vitro test medium. Amorphous The concentration of the thienotriazoloadijephine compound of the formula (1) or the amorphous thienotriazololodiazepine compound (1-1) can be measured at any time during the first 2 hours after the pH adjustment. In some embodiments, the solid dispersion is at least five times as high as the concentration of the polymeric, non-polymeric, thienotriazolol idiazepine compound of formula (1) or crystalline thienothiazololidiazepine compound (1-1) , At least 6-fold higher, at least 7-fold higher, at least 8-fold higher, at least 9-fold higher, or at least 10-fold higher than the amorphous formula (1 ) Of the thienotriazoloadia zepine compound or the amorphous thienotriazololodiazepine compound (1-1).

In another embodiment, a thienotriazololodiazepine compound of amorphous formula (1) or an amorphous thienotriazololadiazine compound of formula (1) from a solid dispersion in an aqueous in vitro test medium having a pH of 1.0 to 2.0, 1) is at least 40%, at least 50%, at least 60%, at least 70%, and at least 50% lower than the concentration of the polymeric, crystalline thienotriazololadiazine compound of formula (1). At least 80% higher. In some of these embodiments, the polymer of the solid dispersion is HPMCAS. In some of these embodiments, the polymer of the solid dispersion is PVP.

In another embodiment, the concentration of the amorphous thienotriazololodiazepine compound of the amorphous formula (1) or the amorphous thienotriazololodiazepine compound (1-1) from the solid dispersion is such that the concentration of the thienotriazole (1) from a solid dispersion of a pharmaceutically acceptable polymer selected from the group consisting of a rhodia zephin compound and a hydromellose phthalate and ethyl acrylate-methyl methacrylate-trimethylammonioethyl methacrylate chloride copolymer. At least 40%, at least 50%, at least 60%, at least 70%, or at least 50%, as compared to the concentration of the thienotriazololodiazepine compound; At least 80% higher, where each solid dispersion was placed in an aqueous test medium in the pH range of 1.0 to 2.0. In some of these embodiments, the polymer of the solid dispersion is HPMCAS. In some of these embodiments, the polymer of the solid dispersion is PVP.

In some embodiments, the solid dispersion described herein exhibits stability to recrystallization of the thienotriazololidiazepine compound (1-1) upon exposure to humidity and temperature over time. In one embodiment, the concentration of the thienotriazololodiazepine compound or thienotriazololodiazepine compound (1-1) of the amorphous formula (1) remaining as amorphous is at least 90%, at least 91%, at least 92% At least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% and at least 99%.

V. Formulation:

Suitable formulations for use with the solid dispersions of the present invention include, without limitation, capsules, tablets, mini tablets, beads, beadlets, pellets, granules, granules, and powders. Suitable formulations can be coated, for example, using enteric-coated skin. Suitable coatings include, without limitation, cellulose acetate phthalate, hydroxypropylmethylcellulose (HPMC), hydroxypropylmethylcellulose phthalate, polymethylacrylic acid copolymer, or hydroxypropylmethylcellulose acetate succinate (HPMCAS). In some embodiments, certain combinations may be encountered, for example, some molecules of the thienothiazololidiazepine of the present invention are present in aggregates while some are in the same sample that is molecularly dispersed with the carrier.

In some embodiments, the solid dispersion of the present invention may be formulated as tablets, caplets, or capsules. In some embodiments, the solid dispersion of the present invention may be formulated as mini-tablets or granules poured into the mouth, or oral powders for constitution. In some embodiments, the solid dispersion of the present invention is dispersed in a suitable diluent in combination with other excipients (e.g., recrystallization / precipitation inhibiting polymers, flavor-masking ingredients, etc.) to provide an instant suspension formulation. In some embodiments, the solid dispersion of the present invention can be formulated for pediatric treatment.

In one embodiment, the pharmaceutical compositions of the present invention are formulated for oral administration. In one embodiment, the pharmaceutical composition is in the form of a solvate, racemate, enantiomer, isomer or isotope labeled form, including a thienotriazolol diazepine compound of formula (1) or a pharmaceutically acceptable salt, hydrate thereof ; And a polymeric carrier. ≪ RTI ID = 0.0 > [0040] < / RTI > In one embodiment, the pharmaceutical composition further comprises at least one additive such as a disintegrant, a lubricant, a lubricant, a binder and a filler.

Examples of pharmaceutically acceptable lubricants and pharmaceutically acceptable lubricants suitable for use in pharmaceutical compositions include, without limitation, colloidal silica, magnesium trisilicate, starch, talc, tribasic calcium phosphate, magnesium stearate, stearic acid Magnesium stearate, aluminum, calcium stearate, magnesium carbonate, magnesium oxide, polyethylene glycol, powdered cellulose, glyceryl behenate, stearic acid, hydrogenated castor oil, glyceryl monostearate, and sodium stearyl fumarate.

Examples of pharmaceutically acceptable binders suitable for use with the pharmaceutical compositions include, without limitation, celluloses and derivatives thereof, such as microcrystalline cellulose (e.g., Avicel PH from FMC), hydroxypropyl cellulose, hydroxyethyl cellulose , And hydroxylpropylmethylcellulose (HPMC, e.g., Methocel of Dow Chemical); Sucrose, dextrose, corn syrup; Polysaccharides; And gelatin.

Examples of pharmaceutically acceptable fillers and pharmaceutically acceptable diluents suitable for use with pharmaceutical compositions include, but are not limited to, sugars, sugars, compressible sugars, dextrates, dextrin, dextrose, lactose, mannitol, microcrystalline cellulose ), Powdered cellulose, sorbitol, sucrose and talc.

In some embodiments, the excipient provides one or more functions in the pharmaceutical composition. For example, the filler or binder may also be a disintegrant, a lubricant, an anti-adhesion agent, a lubricant, a sweetener, and the like.

In some embodiments, the pharmaceutical compositions of this invention may contain additives or components such as antioxidants (e.g., ascorbyl palmitate, butylated hydroxyl anisole (BHA), butylated hydroxytoluene (BHT), alpha (E.g., tocopherol, propyl gallate, and fumaric acid), an antimicrobial agent, an enzyme inhibitor, a stabilizer (e.g., malonic acid), and / or a protective agent.

In general, the pharmaceutical compositions of the present invention may be formulated into any suitable solid dosage form. In some embodiments, the solid dispersion of the present invention is formulated as unit dosage forms for administration, for example as capsules, or tablets, or as multi-particulate systems such as fines or granules or powders.

In one embodiment, the pharmaceutical compositions comprise a solid dispersion of a thienothiazololidiazepine compound of formula (1), and a solid dispersion of hydroxypropylmethyl cellulose acetate succinate (HPMCAS), according to various embodiments of the solid dispersion described herein. , 45-50 wt% lactose monohydrate; 35 to 40% by weight of microcrystalline cellulose; 4-6% by weight of croscarmellose sodium; 0.8-1.5% by weight of colloidal silicon dioxide; And 0.8-1.5 wt.% Magnesium stearate, wherein the thienotriazololidiazepine compound is amorphous in the solid dispersion, and the thienotriazolol diazepine compound versus hydroxypropylmethylcellulose acetate succinate (HPMCAS) The weight ratio is 1: 3 to 1: 1.

VI. Volume:

In one embodiment, the present invention provides a pharmaceutical composition that can be formulated into any suitable solid dosage form. In one embodiment, the pharmaceutical compositions according to the present invention comprise one or more of the various embodiments of the thienothiazololidiazepines of formula (1) as described herein at doses ranging from about 10 mg to about 100 mg do. In one embodiment, the pharmaceutical compositions of the invention comprise from about 10 mg to about 100 mg, from about 10 mg to about 90 mg, from about 10 mg to about 80 mg, from about 10 mg to about 70 mg, from about 10 mg to about 60 mg From about 10 mg to about 50 mg, from about 10 mg to about 40 mg, from about 10 mg to about 30 mg, and from about 10 mg to about 20 mg of a compound of formula (1) as described herein, ) ≪ / RTI > of thienothiazololidiazepines. In one embodiment, the pharmaceutical composition of the present invention is administered at a dose selected from the group consisting of about 10 mg, about 50 mg, about 75 mg, about 100 mg, of a thienotriazololodiazepine of formula And includes one or more of various embodiments.

In one embodiment, the pharmaceutical compositions of the invention comprise about 1 mg, about 2 mg, about 2.5 mg, about 3 mg, about 4 mg, about 5 mg, about 7.5 mg, about 10 mg, about 15 mg, about 20 mg About 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about At a dose selected from the group consisting of about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, and about 150 mg, 1 day for every day, 1 day for every 5 days, 1 day for every 4 days, 1 day for every 3 days, 1 day for every 2 days, 1 time for 1 day, 2 times for 1 time, 1 One or more of the various embodiments of the thienothiazololidiazepines of formula (I) as described herein, in a dosage form selected from the group consisting of three times daily, four times daily, and five times daily. To the subject. In other embodiments, any of the aforementioned dosages or dosage forms are periodically decreased or increased periodically. In one embodiment, the pharmaceutical compositions of the invention comprise about 1 mg, about 2 mg, about 2.5 mg, about 3 mg, about 4 mg, about 5 mg, about 7.5 mg, about 10 mg, about 15 mg, about 20 mg About 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about Wherein the dose is selected from the group consisting of about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, and about 150 mg, Every 1 day for every 5 days, once every day for every 4 days, once every day for every 3 days, once every day for every 2 days, once a day, twice a day, twice a day (1-1), (1-2), (1-3), (1-4), (1-5), ), (1-6), (1-7), (1-8), (1-9), (1-10), (1-11), (1-12) (1-14), (1-15), (1-16), (1-17), and (1-18) to a subject in need thereof Including . In other embodiments, any of the aforementioned dosages or dosage forms are periodically reduced or increased periodically.

Such unit dosage forms are suitable for administration once to five times a day depending on the particular purpose of the therapy, the duration of the therapy, and the like. In one embodiment, the dosage form may be administered to a subject in need thereof at least once a day for at least two consecutive days. In one embodiment, the dosage form can be administered to a subject in need thereof at least once a day, every other day. In one embodiment, the dosage form can be administered to subjects at least weekly in need thereof and can be divided into equivalent and / or unequal doses. In one embodiment, the dosage form may be administered to a subject in need thereof every week, taking into account every 3 days and / or 6 times per week. In one embodiment, the dosage form may be administered to a subject in need thereof every other day, every 3 days, every 4 days, every 5 days, every 6 days and / or every week in divided doses. In one embodiment, the dosage form may be administered to a subject in need of two or more equivalent or unequally divided doses per month.

For example, dosage forms used in capsules, tablets, mini-tablets, beads, beadlets, pellets, granules, granules, or powders may be coated using, for example, enteric-coated tablets. Suitable coatings include, without limitation, cellulose acetate phthalate, hydroxypropylmethylcellulose (HPMC), hydroxypropylmethylcellulose phthalate, polymethylacrylic acid copolymer, or hydroxypropylmethylcellulose acetate succinate (HPMCAS).

VII. Way:

The thienotriazolodiazepine compounds disclosed herein may exist as acid addition salts or as free bases. They can be obtained according to the procedures described in this specification or in U.S. Published Patent Application No. 2010/0286127, which is hereby incorporated by reference in its entirety. The individual enantiomers and diastereoisomers of the thienothiazololidiazepine compounds of the present invention may be prepared synthetically from commercially available starting materials containing asymmetric or stereogenic centers or may be prepared according to methods well known to those skilled in the art ≪ / RTI > or mixtures thereof. Methods of these resolution include: (1) attachment of a mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereoisomers by recrystallization or chromatography, and a glass of optically pure product from the adjuvant, (2) (3) direct separation of a mixture of optical enantiomers on a chiral liquid phase chromatography column, or (4) kinetic partitioning using a stereoselective chemical or enzymatic reagent. Racemic mixtures can also be separated into their component enantiomers by well known methods such as chiral-phase gas chromatography or by crystallization of the compounds in chiral solvents.

If desired, certain enantiomers of thienothiazolazolidinediepine compounds disclosed herein may be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary is cleaved to produce the pure desired Enantiomers are provided. Alternatively, if the molecule contains a basic functional group such as amino or an acidic functional group such as carboxyl, the diastereomeric salt may be formed with a suitable optically active acid or base followed by resolution of the diastereomer, After formation by well-known functional crystallization or chromatography means, the pure enantiomer is recovered. A variety of methods well known in the art can be used to prepare thienotriazoloadia zepine compounds of formula (1) wherein the enantiomeric excess is generally greater than about 80%. Advantageously, the preferred enantiomeric excess is greater than 80%, preferably greater than 90%, more preferably greater than 95%, most preferably 99% and greater.

The solid dispersion of the present invention can be prepared by a number of methods including melting and solvent evaporation. The solid dispersions of the present invention can also be prepared by the methods described in Chiou WL, Riegelman S: "Pharmaceutical applications of solid dispersion systems ", J. Pharm . Sci . 1971; 60: 1281-1302; [Serajuddin ATM: "Solid dispersion of poorly water-soluble drugs: early promises, subsequent problems, and recent breakthroughs ", J. Pharm. Sci. 1999; 88: 1058-1066; [Leuner C, Dressman J: "Improving drug solubility for oral delivery using solid dispersions & Eur . J. Pharm . Biopharm. 2000; 50: 47-60; And [Vasconcelos T, Sarmento B, Costa P: "Solid dispersions as strategy to improve oral bioavailability of poor water soluble drugs", Drug Discovery Today 2007; 12: 1068-1075, all of which are incorporated herein by reference in their entirety.

In one embodiment, the solid dispersion of the present invention is prepared by a melting method. In one embodiment, the melting method comprises melting one or more of the various embodiments of the thienothiazololidiazepine of formula (1) in a carrier. In one embodiment, the melting method comprises cooling the molten compound and the carrier of the present invention. In one embodiment, the fusing process comprises a step of grinding the molten compound and the carrier. In one embodiment, the molten compounds and carrier of the present invention are milled after the cooling step.

In some embodiments, the solvate, racemate, enantiomer, isomer, or isotope labeled form and carrier, including thienotriazolodiazepine of Formula (1) or a pharmaceutically acceptable salt, hydrate thereof, In the case of synthesis, a surfactant is added during the melting step to prevent the formation of suspensions or two liquid layers in the heated mixture. In some embodiments, one or more of the various embodiments of the thienothiazololidiazepine of formula (1) are suspended in a previously molten carrier, instead of using the drug and carrier in the molten state, to lower the process temperature . In one embodiment, the molten drug and carrier mixture is cooled in an ice bath. In one embodiment, the molten drug and carrier mixture is cooled and solidified by spray cooling (alternatively spray coning).

In one embodiment, the melted drug and carrier mixture is sprayed to a cooling chamber through which atmospheric or cooled, low temperature air is passed to form the melt into particles to cool and solidify. In one embodiment, the molten drug and carrier mixture is cooled and solidified by atomization and reconstitution of the molten dispersion in a suitable fluid bed processor. In one embodiment, the molten drug and carrier mixture is cooled and solidified by melt-granulation in a heatable high shear mixer.

In some embodiments, heat sink extrusion or melt agglomeration can be used to avoid melting limits of the drug. Heating device extrusion consists of extrusion at a high speed of the previously mixed drug and carrier at a melting temperature for a short period of time, and the obtained product is recovered after milling at room temperature and milled.

In one embodiment, one or more of the various embodiments of the thienothiazololidiazepines of formula (1) are treated at low processing temperatures to avoid the decomposition of any thermally labile compounds. In one embodiment, a low processing temperature is achieved by associating the extruder with a temporary plasticizer such as carbon dioxide. In one embodiment, melt aggregation is used in the manufacture of solid dispersions according to the present invention in conventional high shear mixers or rotary processors. In one embodiment, the solid dispersion according to the present invention is prepared by adding a molten carrier containing the thienothiazololidiazepine compound according to the invention to a heated excipient. In one embodiment, the solid dispersion according to the present invention is prepared by adding a molten carrier to a heated mixture of a thienotriazolol diazepine and one or more excipients according to the present invention. In one embodiment, the solid dispersion according to the present invention is prepared by heating a mixture of a thienotriazololidiazepine compound according to the present invention, a carrier and one or more excipients at a temperature in the melting range of the carrier or higher.

In some embodiments, one or more of the various embodiments of the formulation of thienotriazoloidazepine according to formula (1) is prepared by a solvent evaporation method. In one embodiment, the solvent evaporation method comprises the step of solubilizing the thienotriazololodiazepine compound according to formula (1) and the carrier in a volatile solvent which is subsequently evaporated. In one embodiment, the volatile solvent may be one or more excipients. In one embodiment, the one or more excipients include, without limitation, an anti-sticking agent, an inert filler, a surfactant wetting agent, a pH adjuster, and an additive. In one embodiment, the excipient may be dissolved, suspended or expanded in a volatile solvent.

In one embodiment, the preparation of a solid dispersion according to the present invention comprises the step of drying one or more excipients suspended in a volatile solvent. In one embodiment, drying includes the use of vacuum drying, slow evaporation of volatile solvents at low temperatures, use of rotary evaporators, spray drying, spray granulation, freeze drying, or supercritical fluid.

In one embodiment, atomization of a suspension or solution of the composition in a small droplet is followed by spray drying of the formulation to a thienotriazololodiazepine composition according to formula (1), including rapid solvent removal from the formulation. In one embodiment, the preparation of a formulation according to the present invention comprises spray granulation in which a solution or suspension of the composition in a solvent is sprayed onto a suitable chemical and / or physical inert filler such as lactose or mannitol. In one embodiment, spray granulation of a solution or suspension of the composition is obtained through a two-way or three-way nozzle.

In some embodiments, the preparation of the solid dispersion according to the present invention involves the use of a supercritical fluid. The term "supercritical fluid" means a material that exists in a single fluid phase above their critical temperature and critical pressure. In one embodiment, the preparation of a formulation according to the present invention involves the use of a supercritical carbon dioxide fluid. In one embodiment, the preparation of a formulation according to the present invention, using supercritical fluid technology, comprises introducing a thienotriazoloidazepine compound according to formula (1) and a carrier simultaneously with carbon dioxide into a particle formation vessel through a nozzle In a conventional solvent; And spraying the solution to allow the solvent to be rapidly extracted by the supercritical fluid so that a precipitate of solid dispersion particles is formed on the vessel wall.

In some embodiments, the preparation of the solid dispersion according to the present invention involves the use of co-precipitation methods. In one embodiment, under constant agitation, the non-solvent is spilled into the thienotriazololodiazepine composition according to formula (1), and the carrier solution. In one embodiment, the thienotriazololodiazepine composition according to formula (1) and the carrier co-precipitate to form a particulate during the addition of the non-solvent. In one embodiment, the resulting particulate is filtered and dried to provide the desired solid dispersion.

The mixing ratio of the compound of formula (1) and the polymorphic carrier (s) is not particularly limited as long as it can improve the bioavailability of the compound of formula (1), and is variable depending on the type of polymer.

The invention is illustrated by the following non-limiting examples.

VIII. Example:

Example 1: In- vitro screening of the solid dispersion of the compound (1-1)

The ten kinds of solid dispersions were prepared by mixing Compound (1-1) and Hypromellose Acetate Succinate (HPMCAS-M), Hypromellose Phthalate (HPMCP-HP55), Polyvinylpyrrolidone (PVP), PVP-Vinyl Acetate PVP-VA), and Yurazit L100-55, with 25% and 50% compound (1-1) loading for each polymer. The solid dispersion was prepared by a solvent evaporation method using spray drying followed by secondary drying in a low temperature convection oven. The performance of each solid dispersion was evaluated by non-impregnation dissolution performance test to measure both the amount of free drug present in the solution and the amount of total drug over time. Non-impregnated dissolution was chosen because it best represents the in vivo situation for low solubility compounds. This test was performed to determine the "uptake" of the dispersion to the intestinal pH (FaNSSIF, pH 6.5) at the stomach pH (0.1N NaCl, pH 1.0) for approximately 30 minutes to 40 minutes after introduction of the dispersion into the test medium, . [FaFSSIF is a section, simulated fasted containing cholate, 0.75 mM lecithin, 0.174 g NaOH _{pellets, 1.977 g NaH 2 PO 4 ·} H 2 O, 3.093 g NaCl, and pure water qs 500 mL in 3 mM sodium tauro fluid. ]. The amount of dissolved drug was quantitated using a high performance liquid chromatography (HPLC) method and an Agilent 1100 series HPLC. The dissolution profiles of the formulations ( Figure 1-Figure 1j ) showed that drug solubility was significantly increased for all of the dispersant candidates compared to the unmethylated compound in the same medium. In a solid dispersion, 25% compound (1-1) in PVP, 25% compound (1-1) in HPMCAS-M, and 50% compound (1-1) in HPMCAS-M release at a long pH Based on the high level of discovery of the free drug, compared to the untreated compound, improved oral absorption was provided.

Example 2: In vivo screening of the solid dispersion of the compound (1-1)

25% Compound (1-1) in PVP, 25% Compound (1-1) in HPMCAS-MG, and 50% Compound (1-1) in HPMCAS-M in the solid dispersion of Compound (1-1) The dispersions were prepared on a larger scale for in vivo experiments. Each formulation was evaluated by the in vitro dissolution test described in Example 1. To ensure that these dispersions are amorphous and homogeneous, each dispersion was evaluated by powder X-ray diffraction (PXRD) and modulated differential scanning calorimetry (mDSC). The X-ray diffractometer was a Bruker D-2 phaser. Additionally, in order to understand the effect of water on the glass transition temperature (Tg) of each dispersion, the mDSC is a sample that was first equilibrated at a set relative humidity (i. E., 25%, 50%, and 75% . Water can act as a plasticizer in the solid dispersion and the hygroscopicity of the system due to the active compound or polymer can affect the water uptake by these systems.

The non-impregnated dissolution results ( FIG. 2-FIG. 2c ) were similar to those found in the dispersion of Example 1. [ The PXRD results ( FIG. 3 ) showed no evidence of crystalline compounds in random dispersions and the mDSC results ( FIGS. 4A- 4C ) showed a single glass transition temperature (Tg) of each dispersion indicating that each dispersion was homogeneous do. An inverse relationship between Tg and relative humidity was observed for each ( Fig. 5 ). Note that for the solid dispersion of 25% compound (1-1) in PVP equilibrated at 75% RH, two Tg appeared, suggesting that phase separation occurred and this dispersion also exhibited a melting event at 75% RH , Suggesting that crystallization occurred during RH equilibration ( FIG. 6 ). This finding implies that the dispersion of 25% compound (1-1) in PVP is less stable than the HPMCAS-M dispersion.

In order to evaluate the bioavailability of these dispersions, an aqueous suspension of a solid dispersion of Compound (1-1) at a dose of 3 mg / kg was administered to a male beagle dog group (3 per group) by oral gavage administration, The compound (1-1) in a mg / kg dose was dissolved in water: ethanol: polyethylene glycol (PEG) 400 (60:20:20) and administered intravenously as a vein in a vein. Blood samples were taken at 0 (before dosing), 5, 15, and 30 minutes and 1, 2, 4, 8, 12, and 24 hours after intravenous administration, and 0 Min, and jugular vein of each animal at 1, 2, 4, 8, 12, and 24 hours. The amount of Compound (1-1) present in each sample was detected using a qualitative LC-MS / MS method with a lower limit value of 0.5 ng / mL. The area under the plasma concentration-time curve (AUC) was determined by using a linear trapezoidal formula up to the last measurable concentration without extrapolation of the terminal isoform until infinity. The elimination half-life (t _1/2 ) was calculated by least squares regression analysis of the terminal linear portion of the log concentration-time curve. Time to peak plasma concentration (C _max) and C _max (t _max) were derived directly from the plasma concentration data. Oral bioavailability (F) was calculated by dividing the dose-normalized AUC after oral administration by the intravenous dose-normalized AUC and recorded as a percentage (%). The results summarized in Table 1 below are the average of the solid dispersions of 25% compound (1-1) in PVP, 25% compound (1-1) in HPMCAS-M and 50% compound (1-1) in HPMCAS-M Oral bioavailability was 58%, 49%, and 74%, respectively.

The pharmacokinetic parameters of the compounds (1-1) after administration of the oral (po) and intravenous (iv) Compound (1-1)
Formulation Capacity & Path C _max
(ng / L) t _max
(hr) AUC
(ng · min / mL) t _1/2
(hr) F (%) Solution in water: ethanol: PEG400 (60: 20: 20) 1 mg / kg
IV 769 0.083 53,312 1.5 ---- 25% water suspension of compound (1-1) / PVP solid dispersion 3 mg / kg
PO 487 1.0 93,271 1.6 58 25% water suspension of compound (1-1) / HPMCAS-M solid dispersion 3 mg / kg
PO 228 0.5 78,595 2.0 49 50% water suspension of compound (1-1) / HPMCAS-M solid dispersion 3 mg / kg PO 371 1.0 118,174 1.5 74 AUC: area under the plasma concentration-time curve; C _max : maximum plasma concentration; F: bioavailability; HPMCAS: sodium hypromellose acetate; IV: intravenous; PEG: polyethylene glycone; PO; Mouth, mouth; PVP: polyvinylpyrrolidone; t _max : time of C _max ; t _1/2 : plasma half-life

Example 3: Preparation and clinical use of capsules containing a solid dispersion of Compound (1-1)

Gelatin capsules at a concentration of 10 mg were prepared for early clinical trials in patients with hematologic malignancies. As described in Examples 1 and 2, on the basis of in vitro and in vivo test results on the solid dispersion of the compound (1-1), 50% of the HPMCAS-M solid dispersion of the compound (1-1) Were selected for capsule development. Capsule development started with a size 3 hard gelatin capsule targeting a loading of 190 mg, because this configuration potentially increases capsule strength by filling larger size capsules while maintaining the pharmaceutical composition. Based on experience, four capsule formulations were designed with different amounts of disintegrant and wetting agent present and absent. The simplest formulations (no wetting agent and minimum disintegration) were chosen for the manufacturing because all four formulations showed similar disintegration and dissolution test results. Manufacturing process development and scale-up experiments were performed to determine the spray drying and drying time for the solid dispersion; Blending parameters; Roller compaction and milling of the blend to obtain a target bulk density of approximately 0.60 g / cc; And capsule filling conditions were confirmed.

Crystalline compound (1-1) and polymeric hydrophromate acetate succinate (HPMCAS-M) were dissolved in acetone and spray dried to produce solid dispersion intermediate (SDI) granules containing 50% compound (1-1) . SDI was identified as amorphous by PXRD analysis and homogeneous by mDSC analysis (i.e., a single Tg under ambient conditions). (1000 g) and microcrystalline cellulose filler-binder (4428 g), croscarmellose sodium disintegrant (636 g), colloidal silicon dioxide dispersant / lubricant (50%) in HPMCAS-M (156 g), magnesium stearate dispersant / lubricant (156 g), and lactose monohydrate filler (5364 g) were blended in the stage of a V-blender. The blend was squeezed and granulated to obtain a bulk density of approximately 0.6 g / mL. The blend was dispensed into a size 3 hard gelatine capsule (target dose: 190 mg) using an automated filling machine and the final capsule was ground with a capsule grinder.

A pharmacokinetic evaluation was performed following oral dosing of 10 mg capsules containing a solid dispersion of 50% Compound (1-1) in HPMCAS and the results were shown in healthy volunteers containing a solid, Compared with pharmacokinetic evaluations performed after oral dosing of 4 x 10 mg capsules.

Comparison results of the two pharmaceutical compositions are provided in Tables 2A and 2B below. The prior Eudragit formulation is described in Example 5 of U.S. Published Patent Application 2009/0012064 A1, published January 8, 2009. This application claims that the preparation of the Eudragit solid dispersion comprises a mixture of the thienotriazoloidazepine of formula (A) and the ammonio methacrylate copolymer type B (Eudragit RS), the methacrylic acid copolymer type C ≪ RTI ID = 0.0 > L100-55), < / RTI > talc and magnesium aluminosilicate were dissolved and / or dispersed in a mixture of water and ethanol. This heterogeneous mixture was then applied to microcrystalline cellulose spheres (Nonpareil 101, Freund) using a centrifugal fluidized bed granulator to produce granules dispensed in size 2 hydroxypropylmethylcellulose capsules.

In both clinical trials, the blood concentration of compound (1-1) was determined using a proven LC-MS / MS method and based on the plasma concentration of compound (1-1) measured at various time points for 24 hours after capsule administration Pharmacokinetic analyzes were performed. The results are summarized in Table 3 below , and the HPMCAS-M solid dispersion formulation was found to have a 3-fold higher bioavailability in humans than the Eudragit solid dispersion formulation based on AUC (924 * 4/1140, administered Adjust for capacity difference). Additionally, based on the observed T _max , the HPMCAS formulation was more rapidly absorbed (T _max = 1 hour versus 4-6 hours) than the Eudragit formulation. A noticeable improvement in systemic exposure to the HPMCAS-M solid dispersion formulation was unexpected.

[Table 2A]

[Table 2B]

After oral administration of the solid dispersion of Compound (1-1) to humans, the pharmacokinetic parameters Compound (1-1) #patient Administration and
Route C _max
(ng / mL) T _max
(hr) AUC _0-24h
(ng · h / mL) Eudragit solid dispersion preparation 7 40 mg PO 83 4 to 6 1140 50% HMPCAS-M solid dispersion preparation 7 10 mg PO 286 One 925 AUC _0-24h : area under the plasma concentration curve of compound (1-1) over time for 24 hours
C _max : maximum concentration in plasma
hr: hour
HPMCAS: Hypromellose acetate succinate
mL: Milliliter
ng: nanogram
PO: mouth, oral
T _max : time of C _max

Example 4. Oral exposure in rats

Oral bioavailability of the three solid preparations of the compound (1-1) was determined in rats. The three selected dispersions were a 25% dispersion of compound (1-1) in PVP, a 25% dispersion of compound (1-1) in HPMCAS-MG and a 50% dispersion of compound (1-1) in HPMCAS- Dispersion. The animal used in the experiment was a specific pathogen-free (SPF) Hsd: Sprague Dawaur rats obtained from the central animal laboratory of the University of Turku, Finland. The rat was originally purchased in Harlan, the Netherlands. The rats were female and 10 weeks old, and 12 rats were used in the experiment. Animals were housed in a polycarbonate Marlboro II (3 animals per us), the room temperature was 21 +/- 3 ° C, the animal room relative humidity was 55 +/- 15% and the animal room brightness was artificially (Cycle of cancer from 18:00 to 06:00) for 12 hours. We used Aspen chips (Tapvei Oy, Estonia) on the bedding and changed the bedding at least once a week. Feed and water were provided before animal dosing but were removed for the first 2 hours after dosing.

A 25% dispersion of compound (1-1) in PVP, a 25% dispersion of compound (1-1) in HPMCAS-MG, and a 50% dispersion of compound (1-1) in HPMCAS- The dosing solution was prepared by adding a pre-calculated amount of injectable sterile water to a container holding the dispersion using an appropriate amount to obtain a compound (1-1) concentration of 0.75 mg / mL. The oral dosing solution was vortexed for 20 seconds before each dose. The intravenous dosing solution containing 0.25 mg / mL of the compound (1-1) contained 4 mL of polyethylene glycol (PEG400) with an average molecular weight of 400 Da, 4 mL of ethanol (96% purity) 5 mg of Compound (1-1) was dissolved in a mixture containing sterile water for use. The dosing solution containing 25% dispersion of compound (1-1) in PVP was used within 30 minutes after water addition. A 25% dispersion of compound (1-1) in HPMCAS-MG and a 50% dispersion of compound (1-1) in HPMCAS-MG were used within 60 minutes after water addition. Dosage levels of compound (1-1) at 1 mg / kg for intravenous administration and 3 mg / kg for oral administration were provided using a dose volume of 4 mL / kg. The dosing schedule is provided in Table 4 below.

Medication plan for rat oral exposure experiment Rat weight Capacity (mL) Test Items Route One 236.5 0.95 Compound (1-1) Intravenous 2 221 0.88 Compound (1-1) Intravenous 3 237.5 0.95 Compound (1-1) Intravenous 4 255.5 1.02 A 25% dispersion of compound (1-1) in PVP oral- 5 224.2 0.90 A 25% dispersion of compound (1-1) in PVP oral- 6 219.2 0.88 A 25% dispersion of compound (1-1) in PVP oral- 7 251.6 1.01 A 25% dispersion of Compound (1-1) in HPMCAS-MG oral- 8 240.4 0.96 A 25% dispersion of Compound (1-1) in HPMCAS-MG oral- 9 238 0.95 A 25% dispersion of Compound (1-1) in HPMCAS-MG oral- 10 226.6 0.91 A 50% dispersion of Compound (1-1) in HPMCAS-MG oral- 11 228.4 0.91 A 50% dispersion of Compound (1-1) in HPMCAS-MG oral- 12 228.5 0.91 A 50% dispersion of Compound (1-1) in HPMCAS-MG oral-

Approximately 50 占 퐇 of blood samples were collected in Eppendorf tubes containing 5 占 퐇 of a solution of ethylenediaminetetraacetic acid (EDTA) at 0.25, 0.5, 1, 2, 4, 8, 12, Were collected within 5 minutes from the specified time point. In each sample, 20 μl of plasma was obtained and stored at dry ice temperature for analysis. Each sample analysis for the concentration of compound (1-1) was performed using a liquid-phase chromatographic tandem mass spectrometry (LC-MS / MS) method validated with a lower limit value of 0.5 ng / mL.

Pharmacokinetic parameters were calculated with the Phoenix WinNonlin software package (version 6.2.1, Pharsight Corp., CA, USA) using the standard non-compartment method. Eliminator half-life (t _1/2 ) was calculated by least squares regression analysis of the end-linear portion of the log concentration-time curve. The area under the plasma concentration-time curve (AUC) was determined by the use of a linear trapezoidal formula up to the last measurable concentration followed by extrapolation of the end remover to infinity. The mean residence time (MRT), which means the mean amount of time remaining in the compartment or system, was calculated by extrapolating the drug concentration profile to infinity. The maximum plasma concentration (C _max) and time (t _max) to the C _max was derived directly from the plasma concentration data. Potential oral bioavailability (F) is calculated by dividing the dose-normalized AUC after oral administration by the dose-normalized AUC after intravenous administration, ie, F = (AUC (oral) / dose (oral)) / (AUC / Dose (intravenous))] and recorded as a percentage (%).

Pharmacokinetic parameters are provided in Table 5 below and graphs of plasma concentration over time are shown in Figures 7 and 8. [

Pharmacokinetic parameters of compound (1-1) after oral and intravenous administration. Values are the mean of three animals. compound parameter 1 mg / kg intravenous 3 mg / kg oral F (%) Compound (1-1) Water: ethanol: PEG400 (60:20:20) AUC (min * ng / mL)
C _max (ng / mL)
T _max (time)
t _1/2 (hour) 8.5
CI / F (mL / min / kg)
MRT (hour) 74698
730
0.25
8.5
13.4
7.4 A 25% dispersion of compound (1-1) in PVP AUC (min * ng / mL)
C _max (ng / mL)
T _max (time)
t _1/2 (hour) 8.5
CI / F (mL / min / kg)
MRT (hour) 39920
77.9
One
13.8
75.2
18.0 18 A 25% dispersion of Compound (1-1) in HPMCAS-MG AUC (min * ng / mL)
C _max (ng / mL)
T _max (time)
t _1/2 (hour) 8.5
CI / F (mL / min / kg)
MRT (hour) 35306
48.3
0.5
11.0
85.0
17.1 16 A 50% dispersion of Compound (1-1) in HPMCAS-MG AUC (min * ng / mL)
C _max (ng / mL)
T _max (time)
t _1/2 (hour) 8.5
CI / F (mL / min / kg)
MRT (hour) 40238
67.0
2
9.5
74.6
12.8 18

Example 5. Preparation of spray-dried dispersion

The spray-dried dispersion of the compound (1-1) was dispersed in a mixture of HPMCAS-MG (Shin Etsu Chemical Co., Ltd.), HPMCP-HP55 (Shin Etsu Chemical Co., Inc., PVP-VA (BASF Corp.), and Eudragit L100-55 (Evonik Industries AG). All spray drying solutions were prepared with 25 wt% and 50 wt% of each polymer. All solutions were prepared in acetone, but the PVP solution was exceptionally prepared in ethanol. For each solution, 1.0 g of a solid (polymer and compound (1-1)) was prepared in 10 g of solvent. The solution was spray dried using a Buchi B-290, PE-024 spray dryer with a nozzle of 1.5 mm and a Buchi B-295, P-002 condenser. The spray dryer nozzle pressure was set at 80 psi, the target discharge temperature was set at 40 ° C, the cooler was set at -20 ° C, the pump speed was set at 100%, and the aspirator setpoint was set at 100%. After spray drying, the solid dispersion was collected and dried overnight in a low-temperature convection oven to remove residual solvent.

Example 6: Stability according to humidity and temperature

The spray-dried dispersion of compound (1-1) in HPMCAS-MG was exposed to moisture at high temperature to evaluate its stability. The glass transition temperature (Tg) according to the relative humidity function was determined at 75% relative humidity, 40 ° C for 1, 2, and 3 months. The spray dried dispersion was stored in an HDPE bottle LDPE bag to simulate bulk product packaging. The results are summarized in Table 6. At 0, Tg was 134 ° C, and at 1 month, Tg was 134 ° C, Tg at 2 months was 135 ° C, and at 3 months Tg was 134 ° C, and only one inflection point was observed at each measurement. An X-ray diffraction pattern was also obtained for each sample. 9 shows the powder X-ray diffraction profile of the solid dispersion of compound (1-1) in HPMCAS-MG at 0 o'clock in the stability test. Figures 10, 11 and 12 show the powder X-ray diffraction patterns of the solid dispersion of compound (1-1) in HPMCAS-MG after 1 month, 2 months and 3 months at 40 ° C and 75% do. This pattern showed no diffraction line associated with compound (1-1).

Example 7: In vitro treatment of triple negative breast cancer cell lines

The inhibitory concentration of compound (1-1) at 50% (GI50) was determined in HCC197, MDA-MB-231 and MDA-MB-468 human TNBC cell lines. Cells were exposed to increasing doses of compound (1-1) for 72 h and cell proliferation was assessed by MTT assay. Growth inhibition 50% (GI50) concentration and maximum effect (Emax) values were calculated using the prism 5.00 for the window and the equation for the S-terminal dose response. Compound (1-1) showed anti-proliferative activity in the cell line three after 72 hours, and the GI50 value was 81.7 to 448.3 nM as shown in Fig.

The results are expressed as a concentration (GI 50) that inhibits 50% of cell growth. Emax% means the maximum inhibitory effect (ratio to control untreated cells) induced by compound (1-1) for cell proliferation. Both values are expressed as an average of 95% confidence intervals. In all cases, n > = 4.

The effect of compound (1-1) on the cell cycle was evaluated after exposure for 24, 48 and 72 hours. Cells were stained with propidium iodide and analyzed using a FACScan flow cytometer. (1-1) (75 nM for MDA-MB-231 cells and 650 nM for two different cell lines) for 24, 48 and 72 hours, respectively, in the HCC1937, MDA-MB-231 and MDA- Respectively. After 72 hours of treatment, the control and treated cells were washed with PBS and re-incubated with the compound (1-1) -free medium (drug wash). 14A-14C, significant differences in the percentage of compound (1-1) -treated cells at the G1, S and G2 / M levels relative to the untreated control group were determined by the one-way ANOVA assay (p <0.001) Followed by the SNK test method. After washing, the significant difference in cell percentage of a given cell cycle phase between the control and pretreated cells was determined by Student's t-test for independent samples, if appropriate, with the same or different variables. *, X, and # indicate the G1 phase (* p <0.05, ** p <0.01), the G2 / M phase (× p <0.05, &Lt; 0.01). &Lt; / RTI &gt; Each bar and vertical line mean mean ± SEM (n ≥ 3), respectively. In the three TNBC cell lines, 24 hr - compound (1-1) exposure significantly increased (p <0.05) the percentage of cells with G1, and cells with S stage decreased. However, in HCC1937 and MDA-MB-231 cells, this blockade was transient because they regained the control cell cycle pattern after 48 hours of drug wash. Interestingly, MDA-MB-468 cell line after drug wash showed a substantial (p &lt; 0.05) accumulation of G2 / M stage cells with loss of G1 stage cells.

Expression levels of BRD2 / 3/4, including c-Myc, were determined by Western blotting and RT-PCR using commercial antibodies at 24, 48, and 72 hours of the three TNBC cell lines, untreated cells and compound (1-1) After exposure, MDA-MB231 cells were analyzed at 75 nM, and HCC1937 and MDA-MB-468 cell lines at 650 nM. RT-PCR was performed using the Fast SYBR Green Master Mix on the Step One Plus real-time PCR system for 24, 48 and 72 hours (panel B). As shown in FIG. 14, significant differences in mRNA levels were determined by the one-tail ANOVA test (p <0.001) followed by the SNK test (* p <0.05, ** p <0.01). Log transformations of the variables were applied before the ANOVA test if necessary. The basal levels of C-MYC, BRD2, BRD3 and BRD4 of HCC1937, MDA-MB-231 and MDA-MB-468 cells are shown in Figures 15A-15E. As shown in Figures 16a-16c, the western blot represents an independent experiment, where each bar and vertical line mean mean ± SEM (n = 3), respectively. As shown in FIGS. 15D-15G, the C-MYC protein and mRNA levels were not changed by treatment with compound (1-1) in all three cell lines. Compound (1-1) down-regulated c-MYC expression (mRNA and protein) in MDA-MB-468 cells (p <0.05). BRD2 levels were increased in MDA-MB231 and MDA-MB-468 cells (p <0.05). Compound (1-1) also decreased the mRNA level of RBD3 in HCC1937 and MDA-MB-468 cells (p < 0.05).

Compound (1-1) was combined with everolimus and the combination index (CI) was determined by the Cho-Talalay method (CI <1, elevation; CI = 1, commercial, CI> 1.1, . The antiproliferative activity of the simultaneous compounds (1-1) and everolimus was evaluated by MTT assay after 72 hours in the indicated cell lines. As shown in FIGS. 17A and 17B, Everolimus had a synergistic effect of synergistic combination with Compound (1-1) in HCC1937 and MDA-MB-231 cells (CI = 1.02 and 0.94, respectively) And antagonistic (CI = 1.60) in MB-468 cell line. The basal expression of BRD2 / 3/4 and C-MYC proteins and mRNA was not correlated with the susceptibility to compound (1-1) or with the combined effect of both drugs.

Example 8: In vivo treatment of mice

In vivo assays were performed in 8-week-old female MDA-MB-231 nude mouse-derived xenografts. When tumor volume reached 100 ㎣, 10 × 10 6 cells were injected and mice were randomly extracted into 4 groups (n = 9): 1) vehicle; 2) 50 mg / kg Compound (1-1) (BID, po); 3) 2 mg / kg of Everolimus (3 times per week, ip); 4) a combination of compound (1-1) and everolimus; Four-week treatment.

As shown in FIG. 18A, tumor weights between different drug plans were compared at each time point during the treatment period and subsequently using the bilateral ANOVA assay (p < 0.001) followed by the Peroney follow-up assay. Symbol ● indicates significant difference between treatments of vehicle treated mice (p <0.05). * Indicates a significant difference (p <0.05, ** p <0.01 and *** p <0.001) in the tumor mass between the combination treated animals and the two single agonist groups. Results are expressed as mean ± SEM (during the treatment period, n = 9). The animal body weight change during treatment is shown in Figure 18B. In vivo, compound (1-1) -treated xenotransplantation significantly reduced tumor mass (p < 0.05) compared to vehicle-treated mice (highest T / C% = 40.7) after 19 days of treatment without weight loss . Everolimus alone was not active, but the combination with compound (1-1) was the most effective treatment strategy (highest T / C% = 20.7). The T / C ratio was 39% after 10 days of combination treatment, and the optimum value was 21% at 23 days. Compound (1-1) can be used alone or in combination with an mTOR inhibitor for the treatment of TNBC.

Example 9

The values of 50% (GI ₅₀ ) for inhibiting the growth of the compound (1-1) were determined by MTT assay and cytometry using 48 hours and 72 hours under normal oxygen and hypoxic (0.1% atmospheric O ₂ ) 231 and MDA-M B-468 human TNBC cell lines. RT-PCR was performed at baseline and at 24, 48, and 72 hours after 500 nM compound (1-1) using the Fast SYBR Green Master Mix in a Step One plus real-time PCR system. Compound (1-1) was combined with docetaxel or mTOR inhibitor, Everolimus and the combination index (CI) was determined by the Chu-Talaray method (CI <0.9, elevation; CI = 0.9-1.1, Respectively.

Compound (1-1) showed antiproliferative activity in three cell lines after treatment for 48 hours and 72 hours under normal oxygen and hypoxic conditions. MDA-MB-231 was the most sensitive in both conditions. Hypoxia significantly increased antiproliferative activity of compound (1-1) in MDA-MB-468 cells ( p <0.05). In addition, the antitumor effect of compound (1-1) was accompanied by a significant ( p < 0.05) reduction in the levels of c-MYC and n-MYC expression in these cells only. 3 cell line showed a substantial increase in p21 expression after compound (1-1) exposure in correlation with G1 cell cycle arrest. Everolimus had adverse effects (CI = 1.02 and 0.94, respectively) in combination with HCC1937 and MDA-MB-231 cell compound (1-1) and antagonistic in MDA-MB-468 cells (CI = 1.60) . Similarly, docetaxel also had a synergistic effect on concurrent combination with compound (1-1) in HCC1937 and MDA-MB-231 cells (CI = 1.03 and 0.95, respectively) and a slight increase in MDA-MB-468 cells CI = 0.87).

These preclinical data supported further development of compound (1-1), either alone or in combination with mTOR inhibitors, in the clinical context for TNBC. A single agent Ib trial is underway in patients with solid tumors including TNBC.

Example 10

The objective of this work was to evaluate the in vitro antitumor activity of compound (1-1) under normal oxygen and hypoxic conditions, including combination with the mTOR inhibitors everolimus and docetaxel.

Experimental procedure: The antiproliferative activity of compound (1-1) was evaluated in three human-derived TNBC cell lines: HCC1937, MDA-MB-231 and MDA-MB-468. Cells were exposed to increased concentrations of compound (1-1) for 48 hours and 72 hours in normal oxygen and hypoxic (0.1% atmospheric O2). MTT assay and cytometry were applied to assess proliferation. The maximum inhibitory effect (Emax%) induced by compound (I-1) at 6 μM and 50% inhibition of cell growth was calculated using the prism 5.00 for the window as an equation for the S- Respectively. The effect of compound (1-1) on the cell cycle was evaluated under normal oxygen conditions after 24, 48 and 72 hours exposure. Cells were stained with propidium iodide and analyzed using a FACScan flow cytometer. As a target of compound (1-1), basic expression levels of BRD2 / 3/4 including c-Myc were analyzed by Western blotting and RT-PCR in TNBC cells under normal oxygen. The mRNA and protein levels of BRD2 / 3/4, c-Myc, n-Myc and CDKN1A (p21) were also assessed after 24, 48 and 72 hours exposure of compound (1-1) at the indicated concentrations in the TNBC cell line model. The antiproliferative effect of compound (1-1) in combination with RAD001 or docetaxel for 48 hours and 72 hours under normal oxygen and hypoxic levels was evaluated in three TNBC cell lines. The combined effect was evaluated using the combination index (CI) determined by the Chu-Talaray method (CI <0.90, increase; 0.90? CI? 1.10, additive effect; CI> 1.10, antagonistic).

As shown in Table 7, the antiproliferative effect of the compound (1-1) was confirmed in HCC1937 and MDA-MB-231 cell lines under normal oxygen and hypoxic conditions. MDA-MB-468 cells were only susceptible to Compound (1-1) under hypoxia.

GI ₅₀ and E _max values of compound (1-1) after 48 hours treatment
Compound (1-1) (48 hours exposure) Normal oxygen Hypoxia Cell line Way GI ₅₀ (nM) E _max % GI ₅₀ (nM) E _max % Cell measurement > 6000 31 > 6000 27 HCC1937 (22-41) (16-39) MTT 109.8 41 23.1 60 (28.6-421.6) (23-59) (8.8-60.6) (54-67) Cell measurement 92.2 71 144.3 67 MDA-MB (63.4-134.0) (64-78) (51.8-396.0) (50-83) MTT 131.9 71 63.3 68 (80.2-217.0) (62-79) (35.3-113.3) (60-76) Cell measurement > 6000 30 60.6 ^* 50 MDA-MB-468 (21-40) (31.3-117.3) (44-56) MTT > 6000 <10 258.0 ^* 53 (80.2-830.1) (31-74)

The results in Table 7 are expressed as a concentration (GI ₅₀ ) that inhibits 50% of cell proliferation. _Emax % represents the maximal inhibitory effect of compound (1-1) on cell proliferation at 6 [mu] M (ratio to control untreated cells). Both values were expressed as an average over a 95% confidence interval. For E _max % ≤ 60%, the IG ₅₀ value was considered to be evident (*). In all cases, n &gt; = 4.

As shown in Fig. 19, in all cell lines, 24 hr-compound (1-1) exposure significantly increased ( p < 0.05) the percentage of cells with G1, while the cells of S stage decreased. HCC1937, MDA-MB-231 and MDA-MB-468 cell lines were simultaneously treated with compound (1-1) for 24, 48 and 72 hours (MDA-MB-231 cells were 75 nM and the other two cell lines were 650 nM). In all cases, cells were stained with propidium iodide and analyzed by flow cytometry. Significant differences in the proportion of G1, S and G2 / M compound (1-1) -treated cells to untreated control cells were determined by SNK post-mortem test following a one-sided ANOVA test (p <0.01). *, ×, and # indicate the G1 cell cycle stage (* p <0.05, ** p <0.01), G2 / M stage (× p <0.05, < / RTI &gt; p < 0.01). Each bar and vertical line represents the mean ± SEM, respectively (n ≥ 3).

As shown in FIGS. 20A and 20B, the compound (1-1) lowered the mRNA level ( p <0.05) of c-Myc expression (mRNA and protein) and BRD4 and n-Myc in MDA-MB- ( P < 0.01). BRD2 levels were increased in MDA-MB-231 and MDA-MB-468 cells ( p <0.05 and p <0.01, respectively). CDKN1A (p21) mRNA levels were increased in all three cell models ( p <0.001). Expression of BRD2 / 3/4, p21, n-Myc and c-Myc at the mRNA and protein levels was determined by RT-PCR and Western blotting on HCC1937, MDA-MB-231 and MDA- 19a) and compound 1-1 exposure (MDA-MB-231 cells were 75 nM and the other two cell lines were 650 nM) for 24, 48 and 72 hours (FIG. 19b). Significant differences in mRNA levels were determined by one-way ANOVA test (p <0.01) followed by SNK test (* p <0.05, ** p <0.01, *** p <0.01). The logarithmic transformation of the variables was applied before the ANOVA test if necessary. Each bar and vertical line represents the mean ± SEM, respectively (n = 3). Western blot represents three independent experiments. (a) n-Myc mRNA was not detected in HCC1937 and MDA-MB-231 cells.

As shown in Table 8, compound (1-1) inhibited proliferation in all TNBC cell lines and had a GI ₅₀ value of < 500 nM after 72 hours of treatment. Similar effects on proliferation were confirmed under cleansing oxygen and hypoxic conditions.

GI ₅₀ and E _max values of compound (1-1) after 72 hours exposure
Compound (1-1) (72 hours exposure) Normal oxygen Hypoxia Cell line Way GI ₅₀ (nM) E _max % GI ₅₀ (nM) E _max % Cell measurement 261.5 70 246.9 54 HCC1937 (38.1-1796) (17-100) (58.1-1049) (31-77) MTT 81.9 ^* 51 29.0 74 (47.0-140.5) (44-58) (2.7-308.6) (54-86) Cell measurement 55.9 89 49.09 82 MDA-MB (45.3-69.0) (84-93) (24.8-96.9) (72-92) MTT 81.7 83 48.2 83 (67.2-99.4) (79-87) (17.1-135.9) (72-95) Cell measurement 303.8 80 251.1 71 MDA-MB-468 (91.5-1008) (50-100) (33.4-1886) (28-100) MTT 448.3 ^* 42 296.2 77 (269.2-746.5) (34-51) (174.2-503.7) (63-91)

The results in Table 8 are expressed as a concentration inhibiting 50% of cell growth (GI ₅₀ ). _Emax % means the maximal inhibitory effect induced by compound (1-1) on cell proliferation at 6 [mu] M (ratio to control untreated cells). Both values were expressed as an average over a 95% confidence interval. If E _max % ≤ 60%, the IG ₅₀ value is considered to be evident (*). In all cases, n &gt; = 4.

Figure 22 shows in vitro evaluation results of the combined effect of Everlloomus (A) or docetaxel (B) and compound (1-1) at 48 hours and 72 hours in TNBC cell lines under normal oxygen and hypoxic conditions. Simultaneous treatment of compound (1-1) with everolimus showed an adverse effect after 72 hours in HCC1937 and MDA-MB-231 cell lines, but was antagonistic in MDA-MB-468 cells under hypoxic and normal oxygen conditions. The combination of compound (1-1) and docetaxel was either additive or slightly syngeneic in TNBC cells. The synergistic effect is defined as CI < 0.9, the CI value of 0.9 to 1.10 means the additive effect of the drug, and CI > 1.10 reflects the antagonism (n = 3).

Conclusion: Compound (1-1) exhibits antiproliferative effect on three types of TNBC cell lines under normal oxygen and hypoxia with a GI 50 value of <500 nM. The growth inhibitory activity of the compound (1-1) was accompanied by a significant accumulation of G1 phase cells, while at the same time the S phase cells decreased and the level of CDKN1A (p21) mRNA increased. The mechanism of action of compound (1-1) appears to be Myc-independent, since the down-regulation of the c-Myc and n-Myc expression occurred only in the MDA-MB-468 cell line. In combination with everolimus, compound (1-1) showed an adverse antiproliferative effect in HCC1937 and MDA-MB-231 cell lines, but was antagonistic in both MDA-MB-468 cells (under both normal and hypoxic conditions). Compounds (1-1) and docetaxel showed an adverse antiproliferative effect in both HCC1937 and MDA-MB-231 cell lines and a synergistic effect in MDA-MB-468 cells in both normal oxygen and hypoxic cells.

Those skilled in the art will appreciate that the illustrative embodiments shown and described above may be varied without departing from the broad inventive concept thereof. Accordingly, it is intended that the present invention not be limited to the illustrative embodiments shown and described, but is intended to cover modifications within the spirit and scope of the invention as defined by the claims. For example, particular features of exemplary embodiments may or may not be part of the claimed invention, and may combine features of the disclosed embodiments. Unless specifically stated otherwise, "one", "one", and "the" are to be understood as meaning "at least one" rather than as an element.

While at least some of the drawings and description of the present invention have been simplified for the sake of clarity in order to focus on components relevant to a clear understanding of the present invention, other components that have been removed for clarity purposes, &Lt; / RTI &gt; However, these components are not well known in the art, and they do not necessarily provide a better understanding of the present invention, so no description of these components is provided herein.

Further, steps in a particular order should not be construed as limitations on the claims, so long as the method is not dependent upon the steps of the particular order described herein. The claims for the method of the present invention should not be limited to the performance of those steps in the order described, and one of ordinary skill in the art will readily understand that the steps are varied and still remain within the spirit and scope of the present invention.

Claims (21)

A method of treating triple negative breast cancer in a mammal comprising administering to a patient a pharmaceutically acceptable amount of a compound, said compound being a thienotriazolol diazepine compound of formula (1) Acceptable salt or hydrate or solvate thereof.

In this formula,
R ¹ is an alkyl carbon number of 1-4,
R ² is a hydrogen atom; A halogen atom; Or alkyl having from 1 to 4 carbon atoms optionally substituted with a halogen atom or a hydroxyl group,
R ³ is a halogen atom; A halogen atom, alkyl having 1 to 4 carbon atoms, alkoxy having 1 to 4 carbon atoms, or cyano; _{^{-NR 5 - (CH 2) m}} -R 6 ( wherein, R ⁵ is a hydrogen atom or an alkyl having 1 to 4 carbons, m is an integer from 0-4, R ⁶ is a halogen atom, an optionally substituted phenyl or flutes Dylem); Or -NR ⁷ -CO- (CH ₂ ) _n -R ⁸ wherein R ⁷ is a hydrogen atom or alkyl having 1 to 4 carbon atoms, n is an integer from 0 to 2, and R ⁸ is optionally substituted with a halogen atom Phenyl or pyridyl,
R ⁴ is - (CH ₂ ) _a -CO-NH-R ⁹ wherein a is an integer of 1 to 4, R ⁹ is alkyl having 1 to 4 carbon atoms, hydroxyalkyl having 1 to 4 carbon atoms, the 1-4 alkoxy; or a 1-4 carbon atoms alkyl, 1-4 carbon atoms alkoxy, amino or hydroxyl group or a phenyl group optionally substituted pyrido dilim) or _{- (CH 2) b -COOR 10} ( wherein b is an integer of 1-4, and R &lt; ¹⁰ &gt; is alkyl having 1-4 carbon atoms. 2. The method of claim 1, further comprising administering at least one chemotherapeutic agent selected from the group consisting of mTOR inhibitors and mitotic inhibitors. 3. The method according to claim 2, wherein the thienotriazoloidazepine compound of formula (1) is administered simultaneously with a chemotherapeutic agent. 3. The method according to claim 2, wherein the thienotriazoloidazepine compound of the formula (1) and the chemotherapeutic agent are administered sequentially. 5. The method according to any one of claims 2 to 4, wherein the chemotherapeutic agent is an mTOR inhibitor. 6. The method of claim 5, wherein the mTOR inhibitor is selected from the group consisting of rapamycin, temsirolimus, lidapololimus, and everolimus. 7. The method of claim 6, wherein the mTOR inhibitor is everolimus. 5. The method according to any one of claims 2 to 4, wherein the chemotherapeutic agent is a mitotic inhibitor. 9. The method of claim 8, wherein the mitotic inhibitor is docetaxel. 10. The compound according to any one of claims 1 to 9, wherein the thienotriazolyldiazepine compound of formula (1)
(i) (S) -2- [4- (4-Chlorophenyl) -2,3,9-trimethyl-6H-thieno [3,2- f] [1,2,4] triazolo- [ , 3-a] [1,4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide or a dihydrate thereof;
(ii) Synthesis of methyl (S) - {4- (3'-cyanobiphenyl-4-yl) -2,3,9-trimethyl-6H- Triazolo [4,3-a] [1,4] diazepin-6-yl} acetate,
(iii) Methyl (S) - (2,3,9-trimethyl-4- (4-phenylaminophenyl) -6H-thieno [3,2- f] [1,2,4] triazolo [ 3-a] [1, 4] diazepin-6-yl} acetate; And
(iv) methyl (S) - {2,3,9-trimethyl-4- [4- (3- phenylpropionylamino) phenyl] -6H- thieno [3,2- f] ] Triazolo [4,3-a] [1,4] diazepin-6-yl} acetate
&Lt; / RTI &gt; 11. The method according to claim 10, wherein the thienotriazolyldiazepine compound represented by formula (1) is (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl-6H-thieno [ 3,2-f] [1,2,4] triazolo- [4,3- a] [1,4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide Way. 11. The method according to claim 10, wherein the thienotriazolyldiazepine compound represented by formula (1) is (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl-6H-thieno [ 3,2-f] [1,2,4] triazolo- [4,3-a] [1,4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide . 13. The therapeutic method according to any one of claims 1 to 12, wherein the thienothiazolazolidiazepine compound is formed as a solid dispersion. 14. The composition of claim 13, wherein the solid dispersion comprises an amorphous thienotriazololadiazepine compound of formula (1), or a pharmaceutically acceptable salt or hydrate thereof; And a pharmaceutically acceptable polymer. 15. The pharmaceutical composition according to claim 14, wherein the pharmaceutically acceptable polymer is hydroxypropylmethyl cellulose acetate succinate (HPMCAS), wherein the weight ratio of thienotriazololodiazepine compound to hydroxypropylmethyl cellulose acetate succinate is from 1: 3 to 1: 1 treatment method. 16. The method according to any one of claims 13 to 15, wherein the solid dispersion exhibits an X-ray powder diffraction pattern substantially free of diffraction lines associated with the crystalline Tienothiazolodiazepine compound of formula (1) Way. 17. The method of any one of claims 13 to 16, wherein the solid dispersion exhibits a single glass transition temperature (Tg) inflection point in the range of from about 130 占 폚 to about 140 占 폚. 18. A process according to any one of claims 13 to 17, wherein the solid dispersion is (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl- -f] [1,2,4] triazolo- [4,3- a] [1,4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide dihydrate A zolodiazepine compound or a pharmaceutically acceptable salt thereof or a hydrate thereof; And a pharmaceutically acceptable polymer. 19. The method of claim 18, wherein the solid dispersion is selected from the group consisting of (S) -2- [4- (4-chlorophenyl) -2,3,9-trimethyl-6H-thieno [ 4] diazepin-6-yl] -N- (4-hydroxyphenyl) acetamide dihydrate, Lt; RTI ID = 0.0 &gt; X-ray powder diffraction pattern. &Lt; / RTI &gt; Claims 1. A compound of formula (1) for use in the treatment of triple-negative breast cancer, or a pharmaceutically acceptable salt or hydrate or solvate thereof:

In this formula,
R ¹ is an alkyl carbon number of 1-4,
R ² is a hydrogen atom; A halogen atom; Or alkyl having from 1 to 4 carbon atoms optionally substituted with a halogen atom or a hydroxyl group,
R ³ is a halogen atom; A halogen atom, alkyl having 1 to 4 carbon atoms, alkoxy having 1 to 4 carbon atoms, or cyano; _{^{-NR 5 - (CH 2) m}} -R 6 ( wherein, R ⁵ is a hydrogen atom or an alkyl having 1 to 4 carbons, m is an integer from 0-4, R ⁶ is a halogen atom, an optionally substituted phenyl or flutes Dylem); Or -NR ⁷ -CO- (CH ₂ ) _n -R ⁸ wherein R ⁷ is a hydrogen atom or alkyl having 1 to 4 carbon atoms, n is an integer from 0 to 2, and R ⁸ is optionally substituted with a halogen atom Phenyl or pyridyl,
R ⁴ is - (CH ₂ ) _a -CO-NH-R ⁹ (wherein a is an integer of 1 to 4, R ⁹ is alkyl having 1 to 4 carbon atoms, hydroxyalkyl having 1 to 4 carbon atoms, the 1-4 alkoxy; or a 1-4 carbon atoms alkyl, 1-4 carbon atoms alkoxy, amino or hydroxyl group or a phenyl group optionally substituted pyrido dilim) or _{- (CH 2) b -COOR 10} ( wherein b is an integer of 1-4, and R &lt; ¹⁰ &gt; is alkyl having 1-4 carbon atoms. A solid dispersion of a compound according to claim 20 and a pharmaceutically acceptable polymer for use in the treatment of triple negative breast cancer.

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